Anti-Galectin-9 antibodies and uses thereof

ABSTRACT

Disclosed herein are anti-Galectin-9 antibodies and methods of using such for inhibiting a signaling pathway mediated by Galectin-9 or eliminating pathologic cells expressing Galectin-9. Such anti-Galectin-9 antibodies may also be used to diagnose and/or to treat diseases associated with Galectin-9, such as autoimmune diseases and solid tumors.

BACKGROUND OF INVENTION

Immune checkpoint blockade has demonstrated unprecedented success in thepast few years as cancer treatment. Often antibodies are used to blockimmune inhibitory pathways, such as the cytotoxicT-lymphocyte-associated protein 4 (CTLA-4) and programmed death 1 (PD-1)pathways. While therapies targeting those two pathways have shownsuccess in treating several cancer types, anti-CTLA-4 and anti-PD-1therapies have a response rate of 10 to 60% of treated patients,depending on cancer type, and have not yet shown the ability to exceed aresponse rate of 60%, even when used in combination (Kyvistborg et al.,Enhancing responses to cancer immunotherapy; Science. 2018 Feb. 2;359(6375):516-517). Additionally, a large number of cancer types arerefractory to these therapies. As part of efforts to improve existingimmunotherapies in the clinic, the field has started to focus on therole of abnormalities in interferon signaling and upregulation ofalternative checkpoints as potential causes for the limitation ofcurrent therapies. One such potential alternate checkpoint is T-cellimmunoglobulin mucin-3 (Tim-3)/Galectin-9 (e.g., reviewed in Yang andHung; The role of T-cell immunoglobulin mucin-3 and its ligandgalectin-9 in antitumor immunity and cancer immunotherapy; Cancerbiology and cancer treatment; October 2017, Vol. 60 No. 10: 1058-1064,and references therein).

Galectin-9 is a tandem-repeat lectin consisting of two carbohydraterecognition domains (CRDs) and was discovered and described for thefirst time in 1997 in patients suffering from Hodgkin's lymphoma (HL)(Tureci et al., J. Biol. Chem. 1997, 272, 6416-6422). Three isoformsexist, and can be located within the cell or extracellularly. ElevatedGalectin-9 levels have been in observed a wide range of cancers,including melanoma, Hodgkin's lymphoma, hepatocellular, pancreatic,gastric, colon and clear cell renal cell cancers (Wdowiak et al. Int. J.Mol. Sci. 2018, 19, 210). In renal cancer, patients with high Galectin-9expression showed more advanced progression of the disease with largertumor size and necrosis (Kawashima et al.; BJU Int. 2014; 113:320-332).In melanoma—a cancer considered as one of the most lethal cancers due toits aggressive metastasis and resistance to therapy—Galectin-9 wasexpressed in 57% of tumors and was significantly increased in the plasmaof patients with advanced melanoma compared to healthy controls (Enningaet al., Melanoma Res. 2016 October; 26(5): 429-441). A number of studieshave shown utility for Gal-9 as a prognostic marker, and more recentlyas a potential new drug target (Enninga et al., 2016; Kawashima et al.BJU Int 2014; 113: 320-332; Kageshita et al., Int J Cancer. 2002 Jun.20; 99(6):809-16, and references therein). Galectin-9 has been describedto play an important role in in a number of cellular processes such asadhesion, cancer cell aggregation, apoptosis, and chemotaxis. Recentstudies have shown a role for Galectin-9 in immune modulation in supportof the tumor, e.g., through negative regulation of Th1 type responses,Th2 polarization and polarization of macrophages to the M2 phenotype.This work also includes studies that have shown that Galectin-9participates in direct inactivation of T cells through interactions withthe T-cell immunoglobulin and mucin protein 3 (TIM-3) receptor(Dardalhon et al., J Immunol., 2010, 185, 1383-1392; Sanchez-Fueyo etal., Nat Immunol., 2003, 4, 1093-1101). Galectin-9 has also been foundto play a role in polarizing T cell differentiation into tumorsuppressive phenotypes), as well as promoting tolerogenic macrophageprogramming and adaptive immune suppression (Daley et al., Nat Med.,2017, 23, 556-567). In mouse models of pancreatic ductal adenocarcinoma(PDA), blockade of the checkpoint interaction between Galectin-9 and thereceptor Dectin-1 found on innate immune cells in the tumormicroenvironment (TME) has been shown to increase anti-tumor immuneresponses in the TME and to slow tumor progression (Daley et al., NatMed., 2017, 23, 556-567). Galectin-9 also has been found to bind toCD206, a surface marker of M2 type macrophages, resulting in a reducedsecretion of CVL22 (MDC), a macrophage derived chemokine which has beenassociated with longer survival and lower recurrence risk in lung cancer(Enninga et al, J Pathol. 2018 August; 245(4):468-477).

Accordingly, modulating the activity of Galectin-9 and/or one or more ofits receptors may provide a novel cancer therapy approach, alone or incombination with existing therapies. Described herein are novel humanantibodies which bind to human Galectin-9 and their therapeutic use inthe treatment of cancer.

SUMMARY OF INVENTION

The present disclosure is based, at least in part, on the development ofanti-Galectin-9 antibodies that potently suppress signaling triggered byGalectin-9. Such antibodies are capable of suppressing Galectin-9signaling and/or eliminating Galectin-9 positive pathologic cells,thereby benefiting treatment of diseases associated with Galectin-9.

Accordingly, one aspect of the present disclosure provides an isolatedanti-Galectin-9 antibody, which binds to an epitope in a carbohydraterecognition domain (CRD) of a Galectin-9 polypeptide, for example, ahuman Galectin-9 polypeptide. In some embodiments, the anti-Galactin-9antibody described herein may bind to both a human Galactin-9polypeptide and a non-human Galactin-9 polypeptide (e.g., a mouseGalactin-9, a rat Galactin-9, or a primate Galactin-9). In someembodiments, the anti-Galactin-9 antibody binds exclusively to one ofthe Galectin-9 CRDs. In some embodiments, the anti-Galactin-9 antibodybinds to both of the Galectin-9 CRDs, e.g., with similar or differentaffinities. In some embodiments, the anti-Galectin-9 antibody disclosedherein binds an epitope within the CRD1 region. In some embodiments, theanti-Galectin-9 antibody disclosed herein binds an epitope within theCRD1 region, which CRD1 region may have the amino acid sequence of SEQID NO: 3. In some embodiments, the anti-Galectin-9 antibody disclosedherein binds an epitope within the CRD1 region having the amino acidsequence of SEQ ID NO: 3. In some embodiments, the anti-Galectin-9antibody binds to the same epitope as a reference antibody selected fromthe group consisting of G9.1-1, G9.1-2, G9.1-3, G9.1-4, G9.1-5, G9.1-6,G9.1-7, G9.1-8, G9.1-8m1, G9.1-8m2, G9.1-8m3, G9.1-8m4, G9.1-8m5,G9.1-8m6, G9.1-8m7, G9.1-8m8, G9.1-8m9, G9.1-8m10, G9.1-8m11, G9.1-8m12,G9.1-8m13, G9.1-8m14, G9.1-9, G9.1-10, and G9.1-11 antibodies, and/orcompetes against the reference antibody from binding to the CRD1 region.In some embodiments, the anti-Galectin-9 antibody binds to the sameepitope as antibody G9.1-8 or antibody G9.1-8m13 and/or competes againstantibody G9.1-8 or antibody G9.1-8m13 from binding to the CRD1 region.

In some embodiments, the anti-Galectin-9 antibody disclosed herein is anantibody selected from the group consisting of G9.1-1, G9.1-2, G9.1-3,G9.1-4, G9.1-5, G9.1-6, G9.1-7, G9.1-8, G9.1-8m1, G9.1-8m2, G9.1-8m3,G9.1-8m4, G9.1-8m5, G9.1-8m6, G9.1-8m7, G9.1-8m8, G9.1-8m9, G9.1-8m10,G9.1-8m11, G9.1-8m12, G9.1-8m13, G9.1-8m14, G9.1-9, G9.1-10, and G9.1-11antibodies. In some embodiments, the anti-Galectin-9 antibody is G9.1-8antibody. In some embodiments, the antibody is G9.1-8m13 antibody. Insome examples, the anti-Galectin-9 antibody may comprise the same heavychain complementarity determining regions (CDRs) and the same lightchain CDRs as the reference antibody, e.g., any of the referenceantibodies provided herein. In one specific example, the anti-Galectin-9antibody comprises the same heavy chain variable region and the samelight chain variable region as the reference antibody, e.g., any of thereference antibodies provided above and elsewhere herein.

In some embodiments, the anti-Galectin-9 antibody has a V_(L) sequencecomprising SEQ ID NO: 21 or consisting essentially of SEQ ID NO: 21 orconsisting of SEQ ID NO: 21. In some embodiments, the anti-Galectin-9antibody has a V_(H) sequence comprising SEQ ID NO: 86 or consistingessentially of SEQ ID NO: 86 or consisting of SEQ ID NO: 86. In someembodiments, the anti-Galectin-9antibody has a V_(L) sequence comprisingSEQ ID NO: 21 and a V_(H) sequence comprising SEQ ID NO: 86. In someembodiments, the anti-Galectin-9antibody has a V_(L) sequence consistingessentially of SEQ ID NO: 21 and a V_(H) sequence consisting essentiallyof SEQ ID NO: 86. In some embodiments, the anti-Galectin-9antibody has aV_(L) sequence consisting of SEQ ID NO: 21 and a V_(H) sequenceconsisting of SEQ ID NO: 86.

In some embodiments, the anti-Galectin antibody has a V_(H) sequencecomprising SEQ ID NO: 22 or consisting essentially of SEQ ID NO: 22 orconsisting of SEQ ID NO: 22. In some embodiments, the anti-Galectin-9antibody has a V_(L) sequence comprising SEQ ID NO: 21 and a V_(H)sequence comprising SEQ ID NO: 22. In some embodiments, theanti-Galectin-9antibody has a V_(L) sequence consisting essentially ofSEQ ID NO: 21 and a V_(H) sequence consisting essentially of SEQ ID NO:22. In some embodiments, the anti-Galectin-9antibody has a V_(L)sequence consisting of SEQ ID NO: 21 and a V_(H) sequence consisting ofSEQ ID NO: 22.

In some embodiments, the anti-Galectin-9 antibody has a V_(L) sequencecomprising one or more of the sequences set forth in SEQ ID NOs: 328,329, and 337. In some embodiments, the anti-Galectin-9 antibody has aV_(H) sequence comprising one or more of the sequences set forth in SEQID NOs: 361, 364, 374, 366, and 383. In some embodiments, theanti-Galectin-9 antibody has a V_(L) sequence comprising one or more ofthe sequences set forth in SEQ ID NOs: 328, 329, and 337, and a V_(H)sequence comprising one or more of the sequences set forth in SEQ IDNOs: 361, 364, and 374. In some embodiments, the anti-Galectin-9antibody has a V_(L) sequence comprising one or more of the sequencesset forth in SEQ ID NOs: 328, 329, and 337, and a V_(H) sequencecomprising one or more of the sequences set forth in SEQ ID NOs: 361,366, and 383.

In some embodiments, the anti-Galectin-9 antibody disclosed herein bindsan epitope within the Galectin-9 CRD2 region. In some embodiments, theanti-Galectin-9 antibody disclosed herein binds an epitope within theGalectin-9 CRD2 region, which CRD2 region may have the amino acidsequence of SEQ ID NO: 4. In some embodiments, the anti-Galectin-9antibody disclosed herein binds an epitope within the CRD2 region havingthe amino acid sequence of SEQ ID NO: 4. In some embodiments, theanti-Galectin-9 antibody binds an epitope within the Galectin-9 CRD2region that comprises a tryptophan residue corresponding with residueW309 of SEQ ID NO: 1. In some embodiments, the anti-Galectin-9 antibodybinds an epitope within the Galectin-9 CRD2 region that does notcomprise one or more residues corresponding with R253, R271, Y330, R334,R341 and Y236 of SEQ ID NO: 1. In some embodiments, the anti-Galectin-9antibody may bind an epitope within the Galectin-9 CRD2 region thatcomprises a tryptophan residue corresponding with residue W309 of SEQ IDNO: 1 and additionally does not comprise one or more residuescorresponding to R253, R271, Y330, R334, R341 and Y236 of SEQ ID NO: 1.In some embodiments, the anti-Galectin-9 antibody binds to the sameepitope as a reference antibody selected from the group consisting ofG9.2-1, G9.2-2, G9.2-3, G9.2-4, G9.2-5, G9.2-6, G9.2-7, G9.2-8, G9.2-9,G9.2-10, G9.2-11, G9.2-12, G9.2-13, G9.2-14, G9.2-15, G9.2-16, G9.2-17,G9.2-17mut6, G9.2-18, G9.2-19, G9.2-20, G9.2-21, G9.2-22, G9.2-23,G9.2-24, G9.2-25, G9.2-26, and G9.2-low affinity binder antibodies,and/or competes against the reference antibody from binding to the CRD2region. In some embodiments, the anti-Galectin-9 antibody binds to thesame epitope as antibody G9.2-17 or antibody G9.2-17mut6 and/or competesagainst antibody G9.2-17 or antibody G92-17mut6 from binding to the CRD2region. In some embodiments, the anti-Galectin-9 antibody is an antibodyselected from the group consisting of G9.2-1, G9.2-2, G9.2-3, G9.2-4,G9.2-5, G9.2-6, G9.2-7, G9.2-8, G9.2-9, G9.2-10, G9.2-11, G9.2-12,G9.2-13, G9.2-14, G9.2-15, G9.2-16, G9.2-17, G9.2-17mut6, G9.2-18,G9.2-19, G9.2-20, G9.2-21, G9.2-22, G9.2-23, G9.2-24, G9.2-25, andG9.2-26 antibodies. In some embodiments, the anti-Galectin-9 antibody isG9.2-17 antibody or G9.2-17mut6 antibody. In some embodiments, theanti-Galectin-9 antibody has a V_(L) sequence comprising SEQ ID NO: 54or consisting essentially of SEQ ID NO: 54 or consisting of SEQ ID NO:54. In some embodiments, the anti-Galectin-9 antibody has a V_(H)sequence comprising SEQ ID NO: 55 or consisting essentially of SEQ IDNO: 55 or consisting of SEQ ID NO: 55. In some embodiments, theanti-Galectin-9antibody has a V_(L) sequence comprising SEQ ID NO: 54and a V_(H) sequence comprising SEQ ID NO: 55. In some embodiments, theanti-Galectin-9antibody has a V_(L) sequence consisting essentially ofSEQ ID NO: 54 and a V_(H) sequence consisting essentially of SEQ ID NO:55. In some embodiments, the anti-Galectin-9antibody has a V_(L)sequence consisting of SEQ ID NO: 54 and a V_(H) sequence consisting ofSEQ ID NO: 55. In some embodiments, the antibody has a V_(H) sequencecomprising SEQ ID NO: 56. In some embodiments, the antibody has a V_(H)sequence consisting essentially of SEQ ID NO: 56 or consisting of SEQ IDNO: 56. In some embodiments, the isolated antibody has a V_(L) sequencecomprising SEQ ID NO: 54 and a V_(H) sequence comprising SEQ ID NO: 56.In some embodiments, the isolated antibody has a V_(L) sequenceconsisting essentially of SEQ ID NO: 54 and a V_(H) sequence consistingessentially of SEQ ID NO: 56. In some embodiments, the isolated antibodyhas a V_(L) sequence consisting of SEQ ID NO: 54 and a V_(H) sequenceconsisting of SEQ ID NO: 56.

In some embodiments, the anti-Galectin-9 antibody has a V_(L) sequencecomprising one or more of the sequences set forth in SEQ ID NOs: 328,329, and 352. In some embodiments, the anti-Galectin-9 antibody has aV_(H) sequence comprising one or more of the sequences set forth in SEQID NOs: 361, 388, 406, and 407. In some embodiments, the anti-Galectin-9antibody has a V_(L) sequence comprising one or more of the sequencesset forth in SEQ ID NOs: 328, 329, and 352, and a V_(H) sequencecomprising one or more of the sequences set forth in SEQ ID NOs: 361,388, and 406. In some embodiments, the anti-Galectin-9 antibody has aV_(L) sequence comprising one or more of the sequences set forth in SEQID NOs: 328, 329, and 352, and a V_(H) sequence comprising one or moreof the sequences set forth in SEQ ID NOs: 361, 388, and 407.

In some examples, the anti-Galectin-9 antibody may comprise the sameheavy chain complementarity determining regions (CDRs) and the samelight chain CDRs as the reference antibody, e.g., any of the referenceantibodies provided herein. In one specific example, the anti-Galectin-9antibody comprises the same heavy chain variable region and the samelight chain variable region as a reference antibody, e.g., any of thereference antibodies provided herein. In some embodiments, theanti-Galectin-9 antibody comprises a heavy chain complementaritydetermining region 1 (CDR1), a heavy chain complementary determiningregion 2 (CDR2), and a heavy chain complementary determining region 3(CDR3), which collectively are at least 90% (e.g., 90%, 91%, 92%, 93%,94%, 95%, 96%, 97%, 98%, or 99%) identical to the heavy chain CDRs of areference antibody, e.g., any of the reference antibodies providedherein. In some embodiments, the anti-Galectin-9 antibody comprises alight chain CDR1, a light chain CDR2, and a light chain CDR3, whichcollectively are at least 90% (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%,97%, 98%, or 99%) identical to the light chain CDRs of a referenceantibody, e.g., any of the reference antibodies provided herein.

In some embodiments, the anti-Galectin-9 antibody comprises both a heavychain complementarity determining region 1 (CDR1), a heavy chaincomplementary determining region 2 (CDR2), and a heavy chaincomplementary determining region 3 (CDR3), which collectively are atleast 90% (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%)identical to the heavy chain CDRs of a reference antibody, e.g., any ofthe reference antibodies provided herein and a light chain CDR1, a lightchain CDR2, and a light chain CDR3, which collectively are at least 90%(e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) identical tothe light chain CDRs of a reference antibody, e.g., any of the referenceantibodies provided herein. In some examples, the anti-Galectin-9antibody may comprise the same heavy chain CDRs and the same light chainCDRs as the reference antibodies noted above. In one specific example,the anti-Galectin-9 antibody may comprise the same heavy chain variableregion and the same light chain variable region as of a referenceantibody, e.g., any of the reference antibodies provided herein. In someembodiments, the exemplary isolated anti-Galectin 9 antibodies whichbind to CRD1 include G9.1-1, G9.1-2, G9.1-3, G9.1-4, G9.1-5, G9.1-6,G9.1-7, G9.1-8, G9.1-9, G9.1-10, G9.1-11, G9.1-8m1, G9.1-8m2, G9.1-8m3,G9.1-8m4, G9.1-8m5, G9.1-8m6, G9.1-8m7, G9.1-8m8, G9.1-8m9, G9.1-8m10,G9.1-8m11, G9.1-8m12, G9.1-8m13, and G9.1-8m14. In some embodiments, theexemplary isolated anti-Galectin 9 antibodies which bind to CRD2 includeG9.2-1, G9.2-2, G9.2-3, G9.2-4, G9.2-5, G9.2-6, G9.2-7, G9.2-8, G9.2-9,G9.2-10, G9.2-11, G9.2-12, G9.2-13, G9.2-14, G9.2-15, G9.2-16, G9.2-17,G9.2-17mut6, G9.2-18, G9.2-19, G9.2-20, G9.2-21, G9.2-22, G9.2-23,G9.2-24, G9.2-25, G9.2-26, and G9.2-low affinity binder.

In some embodiments, the isolated anti-Galectin 9 antibodies, or antigenbinding portion thereof, comprise heavy and light chain variableregions, wherein the light chain variable region comprises an amino acidsequence selected from SEQ ID NO: 29, 13, 34, 36, 38, 40, 42, 44, 46,48, 34, 54, 58, 61, 63, 65, 73, 67, 69, and 71. In some embodiments, thelight chain variable regions consists of an amino acid sequence selectedfrom SEQ ID NO: 29, 13, 34, 36, 38, 40, 42, 44, 46, 48, 34, 54, 58, 61,63, 65, 73, 67, 69, and 71. In some embodiments, the isolatedanti-Galectin 9 antibodies, or antigen binding portions thereof,comprise heavy and light chain variable regions, wherein the heavy chainvariable region comprises an amino acid sequence selected from SEQ IDNO: 30, 31, 32, 33, 35, 37, 39, 41, 43, 45, 47, 49, 50, 51, 52, 53, 55,56, 57, 59, 60, 62, 64, 66, 68, 70, 72 and 73. In some embodiments, theheavy chain variable regions consists of an amino acid sequence selectedfrom SEQ ID NO: 30, 31, 32, 33, 35, 37, 39, 41, 43, 45, 47, 49, 50, 51,52, 53, 55, 56, 57, 59, 60, 62, 64, 66, 68, 70, 72 and 73.

In some embodiments, the isolated anti-Galectin 9 antibodies, or antigenbinding portion thereof, comprise heavy and light chain variableregions, wherein the light chain variable region comprises an amino acidsequence selected from SEQ ID NO: 29, 13, 34, 36, 38, 40, 42, 44, 46,48, 34, 54, 58, 61, 63, 65, 73, 67, 69, and 71, and the heavy chainvariable region comprises an amino acid sequence selected from SEQ IDNO: 30, 31, 32, 33, 35, 37, 39, 41, 43, 45, 47, 49, 50, 51, 52, 53, 55,56, 57, 59, 60, 62, 64, 66, 68, 70, 72 and 73. In some embodiments, theisolated anti-Galectin 9 antibodies, or antigen binding portion thereof,comprise heavy and light chain variable regions, wherein the light chainvariable region consists of an amino acid sequence selected from SEQ IDNO: 29, 13, 34, 36, 38, 40, 42, 44, 46, 48, 34, 54, 58, 61, 63, 65, 73,67, 69, and 71, and the heavy chain variable region consists of an aminoacid sequence selected from SEQ ID NO: 30, 31, 32, 33, 35, 37, 39, 41,43, 45, 47, 49, 50, 51, 52, 53, 55, 56, 57, 59, 60, 62, 64, 66, 68, 70,72 and 73.

In some embodiments, the isolated anti-Galectin 9 antibodies, or antigenbinding portions thereof, comprise heavy and light chain variableregions, wherein the light chain variable region comprises an amino acidsequence selected from SEQ ID NO: 7, 9, 11, 13, 15, 17, 19, 21, 23, 25,and 27. In some embodiments, the light chain variable regions consist ofan amino acid sequence selected from SEQ ID NO: 7, 9, 11, 13, 15, 17,19, 21, 23, 25, and 27. In some embodiments, the isolated anti-Galectin9 antibodies, or antigen binding portions thereof, comprise heavy andlight chain variable regions, wherein the heavy chain variable regioncomprises an amino acid sequence selected from SEQ ID NO: 8, 10, 12, 14,16, 18, 20, 22, 24, 26, 28, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84,85, 86, and 87. In some embodiments, the heavy chain variable regionsconsist of an amino acid sequence selected from SEQ ID NO: 8, 10, 12,14, 16, 18, 20, 22, 24, 26, 28, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83,84, 85, 86, and 87. Accordingly, in some embodiments, provided hereinare isolated anti-Galectin 9 antibodies, or antigen binding portionsthereof, comprising heavy and light chain variable regions, wherein thelight chain variable region comprises an amino acid sequence selectedfrom SEQ ID NO: 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, and 27 and theheavy chain variable region comprises an amino acid sequence selectedfrom SEQ ID NO: 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 74, 75, 76,77, 78, 79, 80, 81, 82, 83, 84, 85, 86, and 87. In some embodiments, thelight chain variable regions consists of an amino acid sequence selectedfrom SEQ ID NO: 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, and 27, and theheavy chain variable regions consists of an amino acid sequence selectedfrom SEQ ID NO: 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 74, 75, 76,77, 78, 79, 80, 81, 82, 83, 84, 85, 86, and 87.

In some embodiments, any of the anti-Galectin-9 antibody disclosedherein may comprise a heavy chain variable domain (V_(H)) that is atleast 85% identical to the V_(H) of a reference antibody disclosedherein. Alternatively or in addition, the anti-Galectin-9 antibody maycomprise a light chain variable domain (V_(L)) that is at least 85%identical to the V_(L) of the reference antibody.

In some embodiments, the anti-Galectin-9 antibody or antigen bindingportion thereof comprises a VL region comprising SEQ ID NO: 54. In someembodiments, the anti-Galectin-9 antibody or antigen binding portionthereof comprises a VH region comprising SEQ ID NO: 55. In someembodiments, the anti-Galectin-9 antibody comprises a VL regionconsisting of SEQ ID NO: 54. In some embodiments, the anti-Galectin-9antibody or antigen binding portion thereof comprises a VH regionconsisting of SEQ ID NO: 55. In some embodiments, the anti-Galectin-9antibody comprises a VL and VH region comprising SEQ ID NO: 54 and 55,respectively. In some specific embodiments, the anti-Galectin-9 antibodyor antigen binding portion thereof comprises a VL and VH regionconsisting of SEQ ID NO: 54 and 55, respectively. In some embodiments,the anti-Galectin-9 antibody is clone 9.2-17. In some embodiments, theanti-Galectin-9 antibody or antigen binding portion thereof comprises aVH region comprising SEQ ID NO: 56. In some embodiments, theanti-Galectin-9 antibody comprises a VL and VH region comprising SEQ IDNO: 54 and 56, respectively. In some specific embodiments, theanti-Galectin-9 antibody or antigen binding portion thereof comprises aVL and VH region consisting of SEQ ID NO: 54 and 56, respectively. Insome embodiments, the anti-Galectin-9 antibody is clone 9.2-17 mut6.

In some embodiments, the anti-Galectin-9 antibody comprises a VL regioncomprising SEQ ID NO: 21. In some embodiments, the anti-Galectin-9antibody comprises a VL region consisting of SEQ ID NO: 21. In someembodiments, the anti-Galectin-9 antibody comprises a VH regioncomprising SEQ ID NO: 86. In some embodiments, the anti-Galectin-9antibody or antigen binding portion thereof comprises a VH regionconsisting of SEQ ID NO: 86. In some embodiments, the anti-Galectin-9antibody comprises a VL and VH region comprising SEQ ID NO: 21 and 86,respectively. In some embodiments, the anti-Galectin-9 antibodycomprises a VL and VH region consisting of SEQ ID NO: 21 and 86,respectively. In some embodiments, the anti-Galectin-9 antibody is cloneG9.1-8m13. In some embodiments, the anti-Galectin-9 antibody comprises aVH region comprising SEQ ID NO: 22. In some embodiments, theanti-Galectin-9 antibody or antigen binding portion thereof comprises aVH region consisting of SEQ ID NO: 22. In some embodiments, theanti-Galectin-9 antibody comprises a VL and VH region comprising SEQ IDNO: 21 and 22, respectively. In some embodiments, the anti-Galectin-9antibody comprises a VL and VH region consisting of SEQ ID NO: 21 and22, respectively. In some embodiments, the anti-Galectin-9 antibody isclone G9.1-8.

In some embodiments, the anti-Galectin-9 antibody or antigen bindingportion thereof comprises a VL region which has the same amino acidsequence as the VL region of antibody 9.1-8m13 (SEQ ID NO: 21). In someembodiments, the anti-Galectin-9 antibody or antigen binding portionthereof comprises a VH region which has the same amino acid sequence asthe VH region of antibody 9.1-8m13 (SEQ ID NO: 86). In some embodiments,the anti-Galectin-9 antibody comprises VL and VH regions which have thesame amino acid sequences as the VL and VH regions of antibody 9.1-8m13(SEQ ID NO: 21 and 86, respectively).

In some embodiments, the anti-Galectin-9 antibody or antigen bindingportion thereof comprises a VL region which has the same amino acidsequence as the VL region of antibody 9.2-17 (SEQ ID NO: 54). In someembodiments, the anti-Galectin-9 antibody or antigen binding portionthereof comprises a VH region which has the same amino acid sequence asthe VH region of antibody 9.2-17 (SEQ ID NO: 55). In some embodiments,the anti-Galectin-9 antibody comprises VL and VH regions which have thesame amino acid sequences as the VL and VH regions of 9.2-17 (SEQ ID NO:54 and 55, respectively).

In some embodiments, the anti-Galectin-9 antibody comprises a VL regionthat has at least 80% (e.g., 85%, 90%, 95%, 96%, 97%, 98%, 99% and anyincrement therein) sequence identity to a VL region set forth in SEQ IDNO: 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, and 27. In some embodiments,the anti-Galectin-9 antibody comprises a VH region that has at least 80%(e.g., 85%, 90%, 95%, 96%, 97%, 98%, 99% and any increment therein)sequence identity to a VH region set forth in SEQ ID NOs: 8, 10, 12, 14,16, 18, 20, 22, 24, 26, 28, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84,85, 86, and 87. In some embodiments, the anti-Galectin-9 antibodycomprises a VL region that has at least 80% (e.g., 85%, 90%, 95%, 96%,97%, 98%, 99% and any increment therein) sequence identity to a VLregion set forth in SEQ ID NOs: 7, 9, 11, 13, 15, 17, 19, 21, 23, 25,and 27 and a VH region that has at least 80% (e.g., 85%, 90%, 95%, 96%,97%, 98%, 99% and any increment therein) sequence identity to a VHregion set forth in SEQ ID NOs: 8, 10, 12, 14, 16, 18, 20, 22, 24, 26,28, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, and 87.

In some embodiments, the anti-Galectin-9 antibody comprises a VL regionthat has at least 80% (e.g., 85%, 90%, 95%, 96%, 97%, 98%, 99% and anyincrement therein) sequence identity to a VL region set forth in SEQ IDNOs: 13, 29, 34, 36, 38, 40, 42, 44, 46, 48, 29, 34, 54, 58, 61, 63, 65,73, 67, 69, and 71. In some embodiments, the anti-Galectin-9 antibodycomprises a VH region that has at least 80% (e.g., 85%, 90%, 95%, 96%,97%, 98%, 99% and any increment therein) sequence identity to a VHregion set forth in SEQ ID NOs: 30, 31, 32, 33, 35, 37, 39, 41, 43, 45,47, 49, 50, 51, 52, 53, 55, 56, 57, 59, 60, 62, 64, 66, 68, 70, 72 and73. In some embodiments, the anti-Galectin-9 antibody comprises a VLregion that has at least 80% (e.g., 85%, 90%, 95%, 96%, 97%, 98%, 99%and any increment therein) sequence identity to a VL region set forth inSEQ ID NOs: 13, 29, 34, 36, 38, 40, 42, 44, 46, 48, 29, 34, 54, 58, 61,63, 65, 73, 67, 69, and 71 and a VH region that has at least 80% (e.g.,85%, 90%, 95%, 96%, 97%, 98%, 99% and any increment therein) sequenceidentity to a VH region set forth in SEQ ID NOs: 30, 31, 32, 33, 35, 37,39, 41, 43, 45, 47, 49, 50, 51, 52, 53, 55, 56, 57, 59, 60, 62, 64, 66,68, 70, 72 and 73.

In some embodiments, the anti-Galectin-9 antibody or antigen bindingportion thereof comprises a VL region that has at least 80% (e.g., 85%,90%, 95%, 96%, 97%, 98%, 99% and any increment therein) sequenceidentity to the VL region set forth in SEQ ID NO: 21.

In some embodiments, the anti-Galectin-9 antibody or antigen bindingportion thereof comprises a VH region that has at least 80% (e.g., 85%,90%, 95%, 96%, 97%, 98%, 99% and any increment therein) sequenceidentity to the VH region set forth in SEQ ID NO: 86. In someembodiments, the anti-Galectin-9 antibody or antigen binding portionthereof comprises a VL and VH region that have at least 80% (e.g., 85%,90%, 95%, 96%, 97%, 98%, 99% and any increment therein) sequenceidentity to the VL and VH regions set forth in SEQ ID NO: 21 and 86,respectively.

In some specific embodiments, the anti-Galectin-9 antibody or antigenbinding fragment thereof comprises a VL that has at least 80% (e.g.,85%, 90%, 95%, 96%, 97%, 98%, 99% and any increment therein) sequenceidentity to a VL region set forth in SEQ ID NO: 54. In some specificembodiments, the anti-Galectin-9 antibody or antigen binding fragmentthereof comprises a VH region that has at least 80% (e.g., 85%, 90%,95%, 96%, 97%, 98%, 99% and any increment therein) sequence identity toa VH region set forth in SEQ ID NO: 55. In some specific embodiments,the anti-Galectin-9 antibody or antigen binding fragment thereofcomprises a VL and/or VH region that has at least 80% (e.g., 85%, 90%,95%, 96%, 97%, 98%, 99% and any increment therein) sequence identity toa VL and/or VH region set forth in SEQ ID NO: 54 and 55, respectively.In some specific embodiments, the anti-Galectin-9 antibody comprises aVL region that has at least 80% (e.g., 85%, 90%, 95%, 96%, 97%, 98%, 99%and any increment therein) sequence identity to the VL region ofG9.1-8m13. In some specific embodiments, the anti-Galectin-9 antibodycomprises a VH region that has at least 80% (e.g., 85%, 90%, 95%, 96%,97%, 98%, 99% and any increment therein) sequence identity to the VHregion of G9.1-8m13. In some specific embodiments, the anti-Galectin-9antibody comprises VL and VH regions that has at least 80% (e.g., 85%,90%, 95%, 96%, 97%, 98%, 99% and any increment therein) sequenceidentity to VL and VH regions of G9.1-8m13. In some specificembodiments, the anti-Galectin-9 antibody comprises a VL region that hasat least 80% (e.g., 85%, 90%, 95%, 96%, 97%, 98%, 99% and any incrementtherein) sequence identity to the VL region of G9.2-17. In some specificembodiments, the anti-Galectin-9 antibody comprises a VH region that hasat least 80% (e.g., 85%, 90%, 95%, 96%, 97%, 98%, 99% and any incrementtherein) sequence identity to the VH region of G9.2-17. In some specificembodiments, the anti-Galectin-9 antibody comprises VL and VH regionsthat has at least 80% (e.g., 85%, 90%, 95%, 96%, 97%, 98%, 99% and anyincrement therein) sequence identity to VL and VH regions of G9.2-17.

Accordingly, in some embodiments, anti-Galectin-9 antibodies or antigenbinding portions thereof comprise (a) VL CDR1 amino acid sequence setforth in SEQ ID NO: 328; (b) VL CDR2 amino acid sequence set forth inSEQ ID NO: 329; (c) VL CDR3 amino acid sequence selected from SEQ ID NO:341-360; (d) VH CDR1 amino acid sequence set forth in SEQ ID NO: 361,427, 428, 431, 435, 436, 437; (d) VH CDR2s amino acid sequence selectedfrom SEQ ID NO: 362, 363, 387-389 and 446-466; (e) VH CDR3 amino acidsequence selected from SEQ ID NO: 390-417. Accordingly, in someembodiments, anti-Galectin-9 antibodies or antigen binding portionsthereof comprise (a) VL CDR1 amino acid sequence set forth in SEQ ID NO:328; (b) VL CDR2 amino acid sequence set forth in SEQ ID NO: 329; (c) VLCDR3 amino acid sequence selected from SEQ ID NO: 330-340; (d) VH CDR1amino acid sequence set forth in SEQ ID NO: 361, 424-434; (e) VH CDR2amino acid sequence selected from SEQ ID NO: 362-366 and 438-445; (f) VHCDR3 amino acid sequence selected from SEQ ID NO: 367-386.

In some embodiments, the anti-Galectin-9 antibody or binding portionthereof comprises heavy and light chain variable regions, wherein thelight chain variable region CDR1 comprises SEQ ID NO: 328. In someembodiments, the anti-Galectin-9 antibody or binding portion thereofcomprises heavy and light chain variable regions, wherein the lightchain variable region CDR2 comprises X₁X₂X₃X₄X₅SX₆X₇X₈SYADSVKG (SEQ IDNO: 467), in which X₁═Y or S, X₂═I or S, X₃═Y or S, X₄═P or S, X₅═Y orS, X₆=G or S, X₇═Y or S, and X₈=T or S. In some embodiments, theanti-Galectin-9 antibody or binding portion thereof comprises heavy andlight chain variable regions, wherein the light chain variable regionCDR3 comprises X₁SX₂X₃X₄X₅X₆X₇X₈X₉X₁₀KX₁₁X₁₂X₁₃GMDY (SEQ ID NO: 468), inwhich X₁═Y or S, X₂=T, S, or absent, X₃═Y, S, or absent, X₄═S or absent,X₅═W, S, or absent, X₆═S or absent, X₇=G, S, or absent, X₈=G, T, S, orabsent, X₉═I, Y, S, or absent, X₁₀=G, S, or Y, X₁₁═W or S, X₁₂═V or S,and X₁₃═W or S. In some embodiments, the anti-Galectin-9 antibody orbinding portion thereof comprises heavy and light chain variableregions, wherein the light chain variable region CDR1 consists of SEQ IDNO: 328. In some embodiments, the anti-Galectin-9 antibody or bindingportion thereof comprises heavy and light chain variable regions,wherein the light chain variable region CDR2 consists ofX₁X₂X₃X₄X₅SX₆X₇X₈SYADSVKG (SEQ ID NO: 467), in which X₁═Y or S, X₂═I orS, X₃═Y or S, X₄═P or S, X₅═Y or S, X₆=G or S, X₇═Y or S, and X₈=T or S.In some embodiments, the anti-Galectin-9 antibody or binding portionthereof comprises heavy and light chain variable regions, wherein thelight chain variable region CDR3 consists ofX₁SX₂X₃X₄X₅X₆X₇X₈X₉X₁₀KX₁₁X₁₂X₁₃GMDY (SEQ ID NO: 468), in which X₁═Y orS, X₂=T, S, or absent, X₃═Y, S, or absent, X₄═S or absent, X₅═W, S, orabsent, X₆═S or absent, X₇=G, S, or absent, X₈=G, T, S, or absent, X₉═I,Y, S, or absent, X₁₀=G, S, or Y, X₁₁═W or S, X₁₂═V or S, and X₁₃═W or S.

In some embodiments, the anti-Galectin-9 antibody or binding portionthereof comprises heavy and light chain variable regions, wherein thelight chain variable region CDR1 comprises SEQ ID NO: 328. In someembodiments, the anti-Galectin-9 antibody or binding portion thereofcomprises heavy and light chain variable regions, wherein the lightchain variable region CDR2 comprises SEQ ID NO: 329. In someembodiments, the anti-Galectin-9 antibody or binding portion thereofcomprises heavy and light chain variable regions, wherein the lightchain variable region CDR3 comprises SEQ ID NO: 337. In someembodiments, the anti-Galectin-9 antibody or binding portion thereofcomprises heavy and light chain variable regions, wherein the lightchain variable region CDR1 consists of SEQ ID NO: 328. In someembodiments, the anti-Galectin-9 antibody or binding portion thereofcomprises heavy and light chain variable regions, wherein the lightchain variable region CDR2 consists of SEQ ID NO: 329. In someembodiments, the anti-Galectin-9 antibody or binding portion thereofcomprises heavy and light chain variable regions, wherein the lightchain variable region CDR3 consists of SEQ ID NO: 337. In someembodiments, the anti-Galectin-9 antibody or binding portion thereofcomprises heavy and light chain variable regions, wherein the lightchain variable region CDR1, CDR2, and CDR3 regions comprise SEQ ID NO:328, 329, and 337, respectively. In some embodiments, the light chainvariable region CDR1, CDR2, and CDR3 regions consist of SEQ ID NO: 328,329, and 337, respectively. In some embodiments, the antibody comprisesthe same VL CDRs as G9.1-8m13. In some embodiments, the anti-Galectin-9antibody or binding portion thereof comprises heavy and light chainvariable regions, wherein the heavy chain variable region CDR1 comprisesSEQ ID NO: 361. In some embodiments, the anti-Galectin-9 antibody orbinding portion thereof comprises heavy and light chain variableregions, wherein the heavy chain variable region CDR2 comprises SEQ IDNO: 366. In some embodiments, the anti-Galectin-9 antibody or bindingportion thereof comprises heavy and light chain variable regions,wherein the heavy chain variable region CDR3 region comprises SEQ ID NO:383. In some embodiments, the anti-Galectin-9 antibody or bindingportion thereof comprises heavy and light chain variable regions,wherein the heavy chain variable region CDR1 consists of SEQ ID NO: 361.In some embodiments, the anti-Galectin-9 antibody or binding portionthereof comprises heavy and light chain variable regions, wherein theheavy chain variable region CDR2 consists of SEQ ID NO: 366. In someembodiments, the anti-Galectin-9 antibody or binding portion thereofcomprises heavy and light chain variable regions, wherein the heavychain variable region CDR3 region consists of SEQ ID NO: 383. In someembodiments, the anti-Galectin-9 antibody or binding portion thereofcomprises heavy and light chain variable regions, wherein the heavychain variable region CDR1, CDR2, and CDR3 regions comprise SEQ ID NO:361, 366, and 383, respectively. In some embodiments, the heavy chainvariable region CDR1, CDR2, and CDR3 regions consist of SEQ ID NO: 361,366, and 383, respectively. In some embodiments, the anti-Galectin-9antibody comprises the same VH CDRs as G9.1-8m13. In one specificembodiment, the anti-Galectin-9 antibody or binding portion thereofcomprises heavy and light chain variable regions, wherein: the lightchain variable region CDR1, CDR2, and CDR3 comprise SEQ ID NO: 328, 329,and 337, respectively, and the heavy chain variable region CDR1, CDR2,and CDR3 comprise SEQ ID NO: 361, 366, and 383, respectively. In someembodiments, the light and heavy chain variable region CDR1, CDR2, andCDR3 regions consist of SEQ ID NO: 328, 329, and 337, respectively, andSEQ ID NO: 361, 366, and 383, respectively. In one specific embodiment,the anti-Galectin-9 antibody comprises the same VL and VH CDRs asG9.1-8m13.

In some embodiments, the anti-Galectin-9 antibody or binding portionthereof comprises heavy and light chain variable regions, wherein thelight chain variable region CDR1 comprises SEQ ID NO: 328. In someembodiments, the anti-Galectin-9 antibody or binding portion thereofcomprises heavy and light chain variable regions, wherein the lightchain variable region CDR2 comprises SEQ ID NO: 329. In someembodiments, the anti-Galectin-9 antibody or binding portion thereofcomprises heavy and light chain variable regions, wherein the lightchain variable region CDR3 comprises SEQ ID NO: 352. In someembodiments, the anti-Galectin-9 antibody or binding portion thereofcomprises heavy and light chain variable regions, wherein the lightchain variable region CDR1 consists of SEQ ID NO: 328. In someembodiments, the anti-Galectin-9 antibody or binding portion thereofcomprises heavy and light chain variable regions, wherein the lightchain variable region CDR2 consists of SEQ ID NO: 329. In someembodiments, the anti-Galectin-9 antibody or binding portion thereofconsists of heavy and light chain variable regions, wherein the lightchain variable region CDR3 comprises SEQ ID NO: 352. In someembodiments, the anti-Galectin-9 antibody or binding portion thereofcomprises heavy and light chain variable regions, wherein the lightchain variable region CDR1, CDR2, and CDR3 regions comprise SEQ ID NO:328, 329, and 352, respectively. In some embodiments, the light chainvariable region CDR1, CDR2, and CDR3 regions consist of SEQ ID NO: 328,329, and 352, respectively. In some embodiments, the anti-Galectin-9antibody comprises the same VL CDRs as G9.2-17. In some embodiments, theanti-Galectin-9 antibody or binding portion thereof comprises heavy andlight chain variable regions, wherein the heavy chain variable regionCDR1, CDR2, and CDR3 regions comprise SEQ ID NO: 361, 388, and 406,respectively. In some embodiments, the heavy chain variable region CDR1,CDR2, and CDR3 regions consist of SEQ ID NO: 361, 388, and 406,respectively. In some embodiments, the antibody comprises the same VHCDRs as G9.2-17. In some embodiments, the anti-Galectin-9 antibody orbinding portion thereof comprises heavy and light chain variableregions, wherein the light chain variable region CDR1, CDR2, and CDR3regions comprise SEQ ID NO: 328, 329, and 352, respectively, and theheavy chain variable region CDR1, CDR2, and CDR3 comprise SEQ ID NO:361, 388, and 406, respectively. In some embodiments, the light andheavy chain variable region CDR1, CDR2, and CDR3 regions consist of SEQID NO: 328, 329, and 352, respectively, and SEQ ID NO: 361, 388, and406, respectively. In one specific embodiment, the anti-Galectin-9antibody comprises the same VL and VH CDRs as G9.2-17.

Accordingly, in some embodiments, anti-Galectin-9 antibodies or antigenbinding portions thereof comprise (a) VL CDR1 amino acid sequence thathas at least 80% (e.g., 85%, 90%, 95%, 96%, 97%, 98%, 99% and anyincrement therein) sequence identity to the VL CDR1 amino acid sequenceset forth in SEQ ID NO: 328; (b) VL CDR2 amino acid sequence that has atleast 80% (e.g., 85%, 90%, 95%, 96%, 97%, 98%, 99% and any incrementtherein) sequence identity to the VL CDR2 amino acid sequence set forthin SEQ ID NO: 329; (c) VL CDR3 amino acid sequence that has at least 80%(e.g., 85%, 90%, 95%, 96%, 97%, 98%, 99% and any increment therein)sequence identity to a VL CDR3 amino acid sequence selected from SEQ IDNO: 330-340; (d) VH CDR1 amino acid sequence that has at least 80%(e.g., 85%, 90%, 95%, 96%, 97%, 98%, 99% and any increment therein)sequence identity to the VH CDR1 amino acid sequence set forth in SEQ IDNO: SEQ ID NO: 361, 427, 428, 431, 435, 436, 437; (e) VH CDR2 amino acidsequence that has at least 80% (e.g., 85%, 90%, 95%, 96%, 97%, 98%, 99%and any increment therein) sequence identity to a VH CDR2 amino acidsequence selected from SEQ ID NO: 362-366 and 438-445; (f) VH CDR3 aminoacid sequence that has at least 80% (e.g., 85%, 90%, 95%, 96%, 97%, 98%,99% and any increment therein) sequence identity to a VH CDR3 amino acidsequence selected from SEQ ID NO: 367-386. Accordingly, in someembodiments, anti-Galectin-9 antibodies or antigen binding portionsthereof comprise (a) VL CDR1 amino acid sequence that has at least 80%(e.g., 85%, 90%, 95%, 96%, 97%, 98%, 99% and any increment therein)sequence identity to the VL CDR1 amino acid sequence set forth in SEQ IDNO: 328; (b) VL CDR2 amino acid sequence that has at least 80% (e.g.,85%, 90%, 95%, 96%, 97%, 98%, 99% and any increment therein) sequenceidentity to the VL CDR2 amino acid sequence set forth in SEQ ID NO: 329;(c) VL CDR3 amino acid sequence that has at least 80% (e.g., 85%, 90%,95%, 96%, 97%, 98%, 99% and any increment therein) sequence identity tothe VL CDR3 amino acid sequence selected from SEQ ID NO: 341-360; (d) VHCDR1 amino acid sequence that has at least 80% (e.g., 85%, 90%, 95%,96%, 97%, 98%, 99% and any increment therein) sequence identity to theVH CDR1 amino acid sequence set forth in SEQ ID NO: 361, 424-434; (d) VHCDR2 amino acid sequence that has at least 80% (e.g., 85%, 90%, 95%,96%, 97%, 98%, 99% and any increment therein) sequence identity to a VHCDR2 amino acid sequence selected from SEQ ID NO: 362, 363, 387-389 and446-466; (e) VH CDR3 amino acid sequence that has at least 80% (e.g.,85%, 90%, 95%, 96%, 97%, 98%, 99% and any increment therein) sequenceidentity to a VH CDR3 amino acid sequence selected from SEQ ID NO:390-417.

In some embodiments, the anti-Galectin-9 antibody or binding portionthereof comprises heavy and light chain variable regions, wherein thelight chain variable region CDR1, CDR2, and CDR3 amino acid sequenceshave at least 80% (e.g., 85%, 90%, 95%, 96%, 97%, 98%, 99% and anyincrement therein) sequence identity to the light chain variable regionCDR1, CDR2, and CDR3 amino acid sequences set forth in SEQ ID NO: 328,329, and 337, respectively. In some embodiments, the antibody VL CDR1,CDR2, and CDR3 amino acid sequences have at least 80% (e.g., 85%, 90%,95%, 96%, 97%, 98%, 99% and any increment therein) sequence identity tothe VL CDR1, CDR2, and CDR3 amino acid sequences of G9.1-8m13. In someembodiments, the anti-Galectin-9 antibody or binding portion thereofcomprises heavy and light chain variable regions, wherein the heavychain variable region CDR1, CDR2, and CDR3 amino acid sequences have atleast 80% (e.g., 85%, 90%, 95%, 96%, 97%, 98%, 99% and any incrementtherein) sequence identity to the heavy chain variable region CDR1,CDR2, and CDR3 amino acid sequences set forth in SEQ ID NO: 361, 366,and 383, respectively. In some embodiments, the antibody VH CDR1, CDR2,and CDR3 amino acid sequences have at least 80% (e.g., 85%, 90%, 95%,96%, 97%, 98%, 99% and any increment therein) sequence identity to theVH CDR1, CDR2, and CDR3 amino acid sequences of G9.1-8m13. In onespecific embodiment, the anti-Galectin-9 antibody or binding portionthereof comprises heavy and light chain variable regions, wherein: thelight chain variable region CDR1, CDR2, and CDR3 amino acid sequenceshave at least 80% (e.g., 85%, 90%, 95%, 96%, 97%, 98%, 99% and anyincrement therein) sequence identity to the light chain variable regionCDR1, CDR2, and CDR3 amino acid sequences set forth in SEQ ID NO: 328,329, and 337, respectively, and the heavy chain variable region CDR1,CDR2, and CDR3 amino acid sequences have at least 80% (e.g., 85%, 90%,95%, 96%, 97%, 98%, 99% and any increment therein) sequence identity tothe heavy chain variable region CDR1, CDR2, and CDR3 amino acidsequences set forth in SEQ ID NO: 361, 366, and 383, respectively. Inone specific embodiment, the antibody VL CDR1, CDR2, and CDR3 and VHCDR1, CDR2, and CDR3 amino acid sequences have at least 80% (e.g., 85%,90%, 95%, 96%, 97%, 98%, 99% and any increment therein) sequenceidentity to the VL CDR1, CDR2, and CDR3 and VH CDR1, CDR2, and CDR3amino acid sequences as G9.1-8m13. In some embodiments, theanti-Galectin-9 antibody or binding portion thereof comprises heavy andlight chain variable regions, wherein the light chain variable regionCDR1, CDR2, and CDR3 amino acid sequences have at least 80% (e.g., 85%,90%, 95%, 96%, 97%, 98%, 99% and any increment therein) sequenceidentity to the light chain variable region CDR1, CDR2, and CDR3 aminoacid sequences set forth in SEQ ID NO: 328, 329, and 352, respectively.In some embodiments, the antibody VL CDR1, CDR2, and CDR3 amino acidsequences have at least 80% (e.g., 85%, 90%, 95%, 96%, 97%, 98%, 99% andany increment therein) sequence identity to the VL CDR1, CDR2, and CDR3amino acid sequences of G9.2-17. In some embodiments, theanti-Galectin-9 antibody or binding portion thereof comprises heavy andlight chain variable regions, wherein the heavy chain variable regionCDR1, CDR2, and CDR3 amino acid sequences have at least 80% (e.g., 85%,90%, 95%, 96%, 97%, 98%, 99% and any increment therein) sequenceidentity to the heavy chain variable region CDR1, CDR2, and CDR3 aminoacid sequences set forth in SEQ ID NO: 361, 388, and 406, respectively.In some embodiments, the antibody VH CDR1, CDR2, and CDR3 amino acidsequences have at least 80% (e.g., 85%, 90%, 95%, 96%, 97%, 98%, 99% andany increment therein) sequence identity to the VH CDR1, CDR2, and CDR3amino acid sequences of G9.2-17. In some embodiments, theanti-Galectin-9 antibody or binding portion thereof comprises heavy andlight chain variable regions, wherein the light chain variable regionCDR1, CDR2, and CDR3 amino acid sequences have at least 80% (e.g., 85%,90%, 95%, 96%, 97%, 98%, 99% and any increment therein) sequenceidentity to the light chain variable region CDR1, CDR2, and CDR3 aminoacid sequences set forth in comprise SEQ ID NO: 328, 329, and 352,respectively, and the heavy chain variable region CDR1, CDR2, and CDR3amino acid sequences have at least 80% (e.g., 85%, 90%, 95%, 96%, 97%,98%, 99% and any increment therein) sequence identity to the heavy chainvariable region CDR1, CDR2, and CDR3 amino acid sequences set forth inSEQ ID NO: 361, 388, and 406, respectively. In one specific embodiment,the antibody VL CDR1, CDR2, and CDR3 and VH CDR1, CDR2, and CDR3 aminoacid sequences have at least 80% (e.g., 85%, 90%, 95%, 96%, 97%, 98%,99% and any increment therein) sequence identity to the VL CDR1, CDR2,and CDR3 and VH CDR1, CDR2, and CDR3 amino acid sequences of G9.2-17.

In some embodiments of any of the anti-Galectin antibodies providedherein, the heavy chain constant region of the anti-Galectin-9 antibodyis from a human IgG (a gamma heavy chain) of any IgG subfamily asdescribed herein, e.g., IgG1 or IgG4.

In some embodiments, the amino acid sequences of exemplary anti-Galectinantibody light chains correspond to sequences set forth in SEQ ID NO:88-98 and SEQ ID NO: 99-115. In some embodiments, the anti-Galectin-9antibodies or antigen-binding portion thereof comprise a light chainsequence of SEQ ID NO: 108. In some embodiments, light chains ofanti-Galectin-9 antibodies comprise an amino acid sequence that has atleast 80% (e.g., 85%, 90%, 95%, 96%, 97%, 98%, 99% and any incrementtherein) sequence identity to the light chain sequence set forth in SEQID NO: 95 (or their variable regions). In some embodiments, light chainsof anti-Galectin-9 antibodies comprise an amino acid sequence that hasat least 80% (e.g., 85%, 90%, 95%, 96%, 97%, 98%, 99% and any incrementtherein) sequence identity to the light chain sequence set forth in SEQID NO: 108 (or their variable regions). In some embodiments, the aminoacid sequences of exemplary anti-Galectin antibody heavy chainscorrespond to sequences set forth in SEQ ID NO: 116-140; 169-193;222-246; 275-299 (anti-Galectin-9 antibodies binding to CRD1) and SEQ IDNO: 141-168; 194-220; 247-274; 300-327 (anti-Galectin-9 antibodiesbinding to CRD2). In some embodiments, the heavy chain constant regionof the anti-Galectin-9 antibody is from a human IgG1. In someembodiments, the anti-Galectin-9 antibodies or antigen-binding portionthereof comprise a heavy chain sequence of SEQ ID NO: 136. In someembodiments, the IgG1 is a mutant with minimal Fc receptor engagement.In some embodiments, the anti-Galectin-9 antibodies or antigen-bindingportion thereof comprise a heavy chain sequence of SEQ ID NO: 189. Insome embodiments, the heavy chain constant region of the anti-Galectin-9antibody is from a human IgG4. In some embodiments, the anti-Galectin-9antibodies or antigen-binding portion thereof comprise a heavy chainsequence of SEQ ID NO: 242. In some embodiments, the IgG4 is IgG4exchange mutant. In some embodiments, the anti-Galectin-9 antibodies orantigen-binding portion thereof comprise a heavy chain sequence of SEQID NO: 295.

In some embodiments, the anti-Galectin-9 antibodies or antigen-bindingportion thereof comprise a heavy chain sequence of SEQ ID NO: 157. Insome embodiments, the anti-Galectin-9 antibodies or antigen-bindingportion thereof comprise a heavy chain sequence of SEQ ID NO: 210. Insome embodiments, the anti-Galectin-9 antibodies or antigen-bindingportion thereof comprise a heavy chain sequence of SEQ ID NO: 263. Insome embodiments, the anti-Galectin-9 antibodies or antigen-bindingportion thereof comprise a heavy chain sequence of SEQ ID NO: 316.

In some embodiments, heavy chains of anti-Galectin-9 antibodies comprisean amino acid sequence that has at least 80% (e.g., 85%, 90%, 95%, 96%,97%, 98%, 99% and any increment therein) sequence identity to the heavychain sequence set forth in SEQ ID NO: 136 (or its variable region). Insome embodiments, heavy chains of anti-Galectin-9 antibodies comprise anamino acid sequence that has at least 80% (e.g., 85%, 90%, 95%, 96%,97%, 98%, 99% and any increment therein) sequence identity to the heavychain sequence set forth in SEQ ID NO: 189 (or its variable region). Insome embodiments, heavy chains of anti-Galectin-9 antibodies comprise anamino acid sequence that has at least 80% (e.g., 85%, 90%, 95%, 96%,97%, 98%, 99% and any increment therein) sequence identity to the heavychain sequence set forth in SEQ ID NO: 242 (or its variable region). Insome embodiments, heavy chains of anti-Galectin-9 antibodies comprise anamino acid sequence that has at least 80% (e.g., 85%, 90%, 95%, 96%,97%, 98%, 99% and any increment therein) sequence identity to the heavychain sequence set forth in SEQ ID NO: 295 (or its variable region). Insome embodiments, heavy chains of anti-Galectin-9 antibodies comprise anamino acid sequence that has at least 80% (e.g., 85%, 90%, 95%, 96%,97%, 98%, 99% and any increment therein) sequence identity to the heavychain sequence set forth in SEQ ID NO: 157 (or its variable region). Insome embodiments, heavy chains of anti-Galectin-9 antibodies comprise anamino acid sequence that has at least 80% (e.g., 85%, 90%, 95%, 96%,97%, 98%, 99% and any increment therein) sequence identity to the heavychain sequence set forth in SEQ ID NO: 210 (or its variable region). Insome embodiments, heavy chains of anti-Galectin-9 antibodies comprise anamino acid sequence that has at least 80% (e.g., 85%, 90%, 95%, 96%,97%, 98%, 99% and any increment therein) sequence identity to the heavychain sequence set forth in SEQ ID NO: 263 (or its variable region). Insome embodiments, heavy chains of anti-Galectin-9 antibodies comprise anamino acid sequence that has at least 80% (e.g., 85%, 90%, 95%, 96%,97%, 98%, 99% and any increment therein) sequence identity to the heavychain sequence set forth in SEQ ID NO: 316 (or its variable region). Insome embodiments, the anti-Galectin-9 antibodies or antigen-bindingportion thereof comprise a light chain sequence of SEQ ID NO: 95 and aheavy chain sequence of SEQ ID NO: 136. In some embodiments, theanti-Galectin-9 antibodies or antigen-binding portion thereof comprise alight chain sequence of SEQ ID NO: 95 and a heavy chain sequence of SEQID NO: 189. In some embodiments, the anti-Galectin-9 antibodies orantigen-binding portion thereof comprise a light chain sequence of SEQID NO: 95 and a heavy chain sequence of SEQ ID NO: 242.

In some embodiments, the anti-Galectin-9 antibodies or antigen-bindingportion thereof comprise a light chain sequence of SEQ ID NO: 95 and aheavy chain sequence of SEQ ID NO: 295. In some embodiments, theanti-Galectin-9 antibodies or antigen-binding portion thereof comprise alight chain sequence of SEQ ID NO: 108 and a heavy chain sequence of SEQID NO: 157.

In some embodiments, the anti-Galectin-9 antibodies or antigen-bindingportion thereof comprise a light chain sequence of SEQ ID NO: 108 and aheavy chain sequence of SEQ ID NO: 210.

In some embodiments, the anti-Galectin-9 antibodies or antigen-bindingportion thereof comprise a light chain sequence of SEQ ID NO: 108 and aheavy chain sequence of SEQ ID NO: 263.

In some embodiments, the anti-Galectin-9 antibodies or antigen-bindingportion thereof comprise a light chain sequence of SEQ ID NO: 108 and aheavy chain sequence of SEQ ID NO: 316.

In one embodiment, the anti-Galectin-9 antibody comprises a light chainamino acid sequence that has at least 80% (e.g., 85%, 90%, 95%, 96%,97%, 98%, 99% and any increment therein) sequence identity to SEQ ID NO:95 and a heavy chain amino acid sequence that has at least 80% (e.g.,85%, 90%, 95%, 96%, 97%, 98%, 99% and any increment therein) sequenceidentity to SEQ ID NO: 136. In one embodiment, the anti-Galectin-9antibody comprises a light chain amino acid sequence that has at least80% (e.g., 85%, 90%, 95%, 96%, 97%, 98%, 99% and any increment therein)sequence identity to SEQ ID NO: 95 and a heavy chain amino acid sequencethat has at least 80% (e.g., 85%, 90%, 95%, 96%, 97%, 98%, 99% and anyincrement therein) sequence identity to SEQ ID NO: 189. In oneembodiment, the anti-Galectin-9 antibody comprises a light chain aminoacid sequence that has at least 80% (e.g., 85%, 90%, 95%, 96%, 97%, 98%,99% and any increment therein) sequence identity to SEQ ID NO: 95 and aheavy chain amino acid sequence that has at least 80% (e.g., 85%, 90%,95%, 96%, 97%, 98%, 99% and any increment therein) sequence identity toSEQ ID NO: 242. In one embodiment, the anti-Galectin-9 antibodycomprises a light chain amino acid sequence that has at least 80% (e.g.,85%, 90%, 95%, 96%, 97%, 98%, 99% and any increment therein) sequenceidentity to SEQ ID NO: 95 and a heavy chain amino acid sequence that hasat least 80% (e.g., 85%, 90%, 95%, 96%, 97%, 98%, 99% and any incrementtherein) sequence identity to SEQ ID NO: 295.

In one embodiment, the anti-Galectin-9 antibody comprises a light chainamino acid sequence that has at least 80% (e.g., 85%, 90%, 95%, 96%,97%, 98%, 99% and any increment therein) sequence identity to SEQ ID NO:108 and a heavy chain amino acid sequence that has at least 80% (e.g.,85%, 90%, 95%, 96%, 97%, 98%, 99% and any increment therein) sequenceidentity to f SEQ ID NO: 157. In one embodiment, the anti-Galectin-9antibody comprises a light chain amino acid sequence that has at least80% (e.g., 85%, 90%, 95%, 96%, 97%, 98%, 99% and any increment therein)sequence identity to SEQ ID NO: 108 and a heavy chain amino acidsequence that has at least 80% (e.g., 85%, 90%, 95%, 96%, 97%, 98%, 99%and any increment therein) sequence identity to SEQ ID NO: 210. In oneembodiment, the anti-Galectin-9 antibody comprises a light chain aminoacid sequence that has at least 80% (e.g., 85%, 90%, 95%, 96%, 97%, 98%,99% and any increment therein) sequence identity to SEQ ID NO: 108 and aheavy chain amino acid sequence that has at least 80% (e.g., 85%, 90%,95%, 96%, 97%, 98%, 99% and any increment therein) sequence identity toSEQ ID NO: 263. In one embodiment, the anti-Galectin-9 antibodycomprises a light chain amino acid sequence that has at least 80% (e.g.,85%, 90%, 95%, 96%, 97%, 98%, 99% and any increment therein) sequenceidentity to SEQ ID NO: 108 and a heavy chain amino acid sequence thathas at least 80% (e.g., 85%, 90%, 95%, 96%, 97%, 98%, 99% and anyincrement therein) sequence identity to SEQ ID NO: 316.

Any of the anti-Galectin-9 antibodies provided herein may comprise aheavy chain variable region framework of VH 3-48; and/or a light chainvariable region framework of Vκ 1-39. In some embodiments, any of the VHand/or VL frameworks described herein are germline VH and/or VL genes.In some embodiments, the anti-Galectin-9 antibodies described herein isa full-length antibody (e.g., an IgG molecule) or an antigen-bindingfragment thereof. In some examples, the antibody is a Fab or asingle-chain antibody. In any instances, the antibody can be a humanantibody or a humanized antibody.

In another aspect, the present disclosure provides an isolated nucleicacid or set of nucleic acids which encode or collectively encode any ofthe anti-Galectin-9 antibodies disclosed herein. In some instances, theheavy chain and light chain of the antibody are encoded by two separatenucleic acid molecules (a set of nucleic acids). In other instances, theheavy chain and light chain of the antibody are encoded by one nucleicacid molecule, which may be in multicistronic format, or under thecontrol of distinct promoters. In some embodiments, the nucleic acid orset of nucleic acids are located on one or two vectors. In someexamples, the one or two vectors can be one or two expression vectors.Further, the present disclosure provides a host cell comprising any ofthe isolated nucleic acid or set of nucleic acids coding for theanti-Galectin-9 antibodies described herein.

Also provided herein is a method for producing the anti-Galectin-9antibody, comprising culturing the host cell described herein undersuitable conditions allowing for expressing of the antibody, andharvesting the antibody thus produced from the cell culture (e.g., fromthe culture medium).

Further, the present disclosure provides a pharmaceutical composition,comprising any of the anti-Galectin-9 antibodies or a nucleic acid(s)encoding such, and a pharmaceutically acceptable carrier.

In yet another aspect, the present disclosure features a method ofinhibiting Galectin-9-mediated cell signaling in a subject, the methodcomprising administering to a subject in need thereof an effectiveamount of an anti-Galectin-9 antibody or a pharmaceutical compositioncomprising an anti-Galectin-9 antibody. In some embodiments, theanti-Galectin-9 antibody is any of the anti-Galectin-9 antibodiesdisclosed herein or a pharmaceutical composition comprising such. Insome embodiments, the subject in need thereof is a human patient having,suspected of having, or at risk for having, an autoimmune disease, asolid cancer, a microbial disease, a hematological malignancy, or anallergic disorder. Exemplary autoimmune diseases include, but are notlimited to, a rheumatoid condition (e.g., rheumatoid arthritis), anautoimmune respiratory disease, an autoimmune metabolic and/or endocrinedisorder (e.g., type I diabetes), or a fibrotic condition. Exemplarysolid tumors include, but are not limited to, pancreatic ductaladenocarcinoma (PDA), colorectal cancer (CRC), melanoma,cholangiocarcinoma, breast cancer, small cell and non small cell lungcancer, upper and lower gastrointestinal malignancies, gastric cancer,squamous cell head and neck cancer, genitourinary cancer, hepatocellularcarcinoma, ovarian cancer, sarcomas, mesothelioma, glioblastoma,esophageal cancer, bladder cancer, urothelial cancer, renal cancer,cervical and endometrial cancer. Exemplary hematological malignanciesinclude, but are not limited to, acute lymphoblastic leukemia, chroniclymphocytic leukemia, lymphomas, multiple myeloma, and acute myelogenousleukemia, chronic myelogenous leukemia, myelodysplastic syndromes, ormyeloproliferative neoplasms and other myeloproliferative andmyelodysplastic disorders. In some examples, the effective amount of thepharmaceutical composition is sufficient to block interaction betweenGalectin-9 and Dectin-1. In some embodiments, the effective amount ofthe pharmaceutical composition is sufficient to block interactionbetween Galectin-9 and CD206. Alternatively, or in addition, but notlimited to, the effective amount of the pharmaceutical composition issufficient to block interaction between Galactin-9 and Tim-3.

Further, the present disclosure provides a method for modifying,eliminating and/or reducing pathologic cells expressing Galectin-9(e.g., via antibody-dependent cell cytotoxicity or ADCC), the methodcomprising administering to a subject having pathologic cells expressingGalectin-9 an effective amount of an anti-Galectin-9 antibody, such asany of the anti-Galectin-9 antibodies described herein, or apharmaceutical composition thereof. In some embodiments, the subject isa human patient having cancer cells expressing Galectin-9 and/orpathologic immune cells expressing Galectin-9. In some embodiments, theeffective amount of the pharmaceutical composition is sufficient toinitiate antibody-dependent cell cytotoxicity (ADCC) and/or blockagainst pathologic cells expressing Galectin-9.

Any of the treatment methods described herein may further compriseadministering to the subject an inhibitor of a checkpoint molecule, anactivator of a co-stimulatory receptor, or an inhibitor of an innateimmune cell target. Examples of checkpoint molecules include, but arenot limited to, PD-1, PD-L1, PD-L2, CTLA-4, LAG3, TIM-3 and A2aR.Examples of co-stimulatory receptors include, but are not limited to,OX40, GITR, CD137, CD40, CD27, and ICOS. Examples of innate immune celltargets include, but are not limited to, KIR, NKG2A, CD96, TLR, and IDO.

The present disclosure also provides pharmaceutical compositions for usein treating a disease associated with Galectin-9 (e.g., those describedherein), wherein the pharmaceutical composition comprises ananti-Galectin-9 antibody, such as any of the anti-Galectin-9 antibodiesdescribed herein, or a nucleic acid(s) encoding such antibody, and apharmaceutically acceptable carrier. Also, the present disclosureprovides uses of the anti-Galectin-9 antibodies or the encoding nucleicacids for manufacturing a medicament for use in treating the targetdiseases as described herein.

The details of one or more embodiments of the invention are set forth inthe description below. Other features or advantages of the presentinvention will be apparent from the following drawing and detaileddescription of several embodiments, and also from the appended claims.

BRIEF DESCRIPTION OF DRAWINGS

The following drawings form part of the present specification and areincluded to further demonstrate certain aspects of the presentdisclosure, which can be better understood by reference to the drawingin combination with the detailed description of specific embodimentspresented herein.

FIGS. 1A-1B include charts showing a binding characterization of Fabsfor Galectin-9 CRD2 using phage ELISAs. FIG. 1A: binding to human andmouse Galectin-9 shown by phage ELISA. FIG. 1B: affinity of Fabs clonesto Galectin-9 CRD2 determined by competition phage ELISA.

FIGS. 2A-2B include charts showing a binding characterization of Fabsfor Galectin-9 CRD1 using phage ELISA. FIG. 2A: binding of Fab clones tohuman and mouse Galectin-9 CRD1 shown by phage ELISA. FIG. 2B: affinityof Fabs clones to Galectin-9 CRD1 determined by competition phage ELISA.

FIGS. 3A-3B include charts showing epitope binning of G.9-2 Fab clones(binding to CRD2) using competition phage ELISA. FIG. 3A: mouseGalectin-9 CRD2-coated wells pre-incubated with purified G9.2-1 orG9.2-3 Fabs prior to addition of phage-displayed Galectin-9 CRD2 bindingFab clones. FIG. 3B: human Galectin-9 CRD2-coated wells pre-incubatedwith purified G9.2-15 or G9.2-17 Fabs prior to addition ofphage-displayed Galectin-9 CRD2 binding Fab clones.

FIG. 4 includes diagrams showing the affinity of purified G9.2 Fabs toGalectin-9 CRD2, characterized using a bead-based binding assay. Thecurves show the best fit of the one-to-one binding model. Top left:G9.2-1 Fab. Top right: G9.2-3 Fab. Bottom left: G9.2-15 Fab. Bottomright: G9.2-17 Fab. Apparent Kd values are shown in the table.

FIG. 5 includes diagrams showing the affinity of purified G9.1 Fabs toGalectin-9 CRD1, characterized using a bead-based binding assay.Experiments were performed in the same manner as in FIG. 4. Top left:G9.1-6 Fab. Top right: G9.1-5 Fab. Bottom left: G9.1-8 Fab. Bottomright: G9.1-11 Fab. Apparent Kd values are shown in the table.

FIG. 6 includes diagrams showing a surface plasmon resonance analysis ofFab G9.2-15 and Fab G9.2.17 binding to CRD2 of human (top) and mouse(bottom) Galectin-9. The binding and dissociation phases of theexperiments are marked in the top panels. Left: G9.2-15 Fab. Right:G9.2-17 Fab.

FIG. 7 includes diagrams showing an SPR analysis of G9.2-17 human IgG4binding to CRD2 of human (top) and mouse (bottom) Galectin-9. The graylines show the sensorgrams for the non-binding negative control,G9.2-iso human IgG4.

FIG. 8 includes diagrams showing the staining of cell line samples withFabs for Galectin-9 CRD2. Histograms for flow cytometry data are shown.Top left: G9.2-1 Fab. Top right: G9.2-3 Fab. Bottom left: G9.2-15 Fab.Bottom right: G9.2-17 Fab.

FIG. 9 is a chart showing the inhibitory effects of G9.2-17 and G9.1-8on Galectin-9 mediated activation of Dectin-1 signaling.

FIGS. 10A-10B include diagrams showing epitope mapping of G.9-2.17 onhuman Galectin-9 CRD2 by systematic mutagenesis. FIG. 10A: A diagramshowing the binding activity of G9.2-17 to Galectin-9 CRD2 mutants asdetermined by phage ELISA. The reduction in ELISA signal indicates asite on the Galectin-9 CRD2 that is critical to G9.2-17 binding. FIG.10B: a diagram depicting the location of W309 as mapped on the crystalstructure of human Galectin-9 CRD2 (PDB ID 3NV2), which is opposite tothe binding site of the sugar ligand as mapped on the crystal structure(W309 corresponds with W277 in UniProt ID 000182-2; PDB ID 3NV2).

FIG. 11 contains charts showing size-exclusion chromatography analysesof Fab G9.2-17 (top), Fab G9.2-17mut6 (middle) and Fab G9.2-Iso(bottom). Purified Fab samples were run on TOSOH TSK Bioassist G2WXLColumn in PBS and detected using absorbance at 280 nm.

FIG. 12 contains charts showing surface plasmon resonance analyses ofFab G9.2-17 (top) and Fab G9.2.17mut6 (bottom) binding to the CRD2 ofhuman (left) and mouse (right) Galectin-9. Human or mouse Galectin-9CRD2 was immobilized on an Avicap chip preloaded with neutravidin on aPall ForteBio Pioneer instrument. Fab samples were then flowed using theOneStep method. The binding and dissociation phases of the experimentsare marked in the top panels.

FIG. 13 is a graph showing a binding characterization of G9.2 Fab clonefor wild-type Galectin-9 CRD2 or the W3039K mutant using phage ELISA.Binding of Fab clones to human Galectin-9 CRD2 assayed using phageELISA. Either biotinylated wild type human Galectin-9 CRD2, the W309KGalectin-9 CRD2 mutant, or Galectin-9 CRD2 pre-incubated with G9.2-17IgG was immobilized to neutravidin-coated wells and incubated withindividual phage-displayed Fab clones.

FIG. 14 is a Kaplan-Meier plot showing that blocking Galectin-9 resultsin significant extension of survival in animal models of pancreaticcancer (KPC mice).

FIG. 15 is a photograph of mouse tumors showing that blocking galectin-9and anti-PD1 generates a superior response.

FIG. 16 is a bar graph showing the tumor mass of mice treated withG9.2-17 mIgG1. Mice (n=10/group) with orthotopically implanted KPCtumors were treated with commercial isotype (200 μg) or commercial αGal9(200 μg) mAb or G9.2-Iso mIgG1 (200 μg) or G9.2-17 mIgG1 at two doses(200 μg or 400 μg) once weekly for three weeks. Tumors were removed andweighed, and subsequently processed and stained for flow cytometry.

FIG. 17 depicts a bar graph showing tumor weight of mice treated withG9.2-17 mIgG2a alone or in combination with αPD1 mAb. Mice (n=10/group)with orthotopically implanted KPC tumors were treated with commercialαPD-1 (200 μg) mAb or G9.2-17 mIg2a (200 μg), or a combination ofG9.2-17 and αPD-1, or matched isotype once weekly for three weeks.Tumors were removed and weighed and subsequently processed and stainedfor flow cytometry. Each point represents one mouse; *p<0.05; **p<0.01;***p<0.001; ****p<0.0001; by unpaired Student's t-test.

FIGS. 18A-18C depict graphs showing binding of purified G9.1-8m1-5 mIgG1to human Galectin-9 CRD1 as characterized using a bead-based bindingassay.

FIGS. 19A-19G depict graphs showing binding of purified G9.1-8m6-11 Fabsto human Galectin-9 CRD1 as characterized using a bead-based bindingassay.

FIGS. 20A-20C depict graphs showing the affinity of purified G9.1-8m8,9, and 11 mIgG2a antibodies to human Galectin-9 CRD1 as characterizedusing a bead-based binding assay.

FIGS. 21A-21D depict graphs showing binding of purified G9.1-8m11-14Fabs to human Galectin-9 CRD1 as characterized using a bead-basedbinding assay.

FIGS. 22A-22D depict graphs showing binding of purified G9.1-8m12-14mIgG2a antibodies to human Galectin-9 CRD1 as characterized using abead-based binding assay.

FIG. 23 depicts a graph showing the results of an apoptosis assaydemonstrating that Gal-9 antibodies inhibit Galectin-9 induced apoptosisof Jurkat cells. Jurkat cells were treated with or without Galectin-9(280 nM), G9.2-17 IgG (1 and/or G9.1-8m13 IgG (1 μM) for 6 hours. Cellswere then stained with annexin-V and PI followed by flow cytometryanalysis. AnnexinV positive cells represent cells in both early and latestage apoptosis. Bars represent average of three replicates, representedas individual data points. Statistical analysis performed by unpairedStudent's t-test. (*p<0.05; **p<0.01; ***p<0.001; ****p<0.0001).

FIG. 24 depicts a graph showing the readout of assays demonstratinganti-Galectin-9 antibodies disclosed herein disrupt the interactionbetween Galectin-9 and CD206. FIG. 24A depicts a graph showing an ELISAmeasuring the interaction between immobilized human Galectin-9 andsoluble CD206 in the absence and presence of the addition of G9.1-8m13,or G9.2-17 antibody. Isotype antibody wells serves as control.Galectin-9 coated wells were incubated with CD206 with or withoutG9.1-8m13, G9.2-17, a combination of both antibodies, or an isotype.(Experiments performed in triplicate; *p<0.05; **p<0.01; ***p<0.001;****p<0.0001; by unpaired Student's t-test). These results indicate thatboth G9.1-8m13 and G9.2-17 antibodies inhibit the interaction betweenGalectin-9 and CD206 and their effects are additive.

FIG. 25 depicts a line graph showing binding of purified G9.1-8m12-14mIgG2a antibodies to human Galectin-9 CRD2 as compared to G9.18 (WT) ascharacterized using a bead-based binding assay.

FIGS. 26A and 26B depict bar graphs showing TNF-alpha (FIG. 26A) andIFNgamma (FIG. 26B) expression in CD3+ T cells in pancreaticadenocarcinoma primary tumor sample patient-derived organotypic tumorspheroids (PDOTs) treated with 9.2-17 IgG4 (100 nM) as compared toisotype control (100 nM).

FIGS. 27A-27C depict bar graphs showing CD44 (FIG. 27A), TNF-alpha (FIG.27B) and IFNgamma (FIG. 27C) expression in CD3+ T cells in pancreaticadenocarcinoma primary tumor sample patient-derived organotypic tumorspheroids (PDOTS) treated with 9.2-17 IgG1 (100 nM) or 9.2-17 IgG4 (100nM) as compared to IgG1 or IgG4 isotype control (100 nM).

FIGS. 28A-28F depict bar graphs showing immune profile expression in aGall Bladder Cancer tumor sample (PDOTS) treated with 9.2-17 IgG4 (100nM) as compared to IgG4 isotype control (100 nM); CD44 in CD3+ T cells(FIG. 28A), TNF-alpha in CD3+ T cells (FIG. 28B), CD44 in CD4+ T cells(FIG. 28C), TNF-alpha in CD4+ T cells (FIG. 28D), CD44 in CD8+ T cells(FIG. 28E), TNF-alpha in CD8+ T cells (FIG. 28F).

FIGS. 29A-29C depict bar graphs showing CD44 (FIG. 29A), TNF-alpha (FIG.29B) and IFNgamma (FIG. 29C) expression in CD3+ T cells in a sample ofliver metastasis from a colorectal cancer patient (PDOTs) treated with9.2-17 IgG1 (100 nM) or 9.2-17 IgG4 (100 nM) as compared to IgG1 (100nM) or untreated control (Utx).

FIG. 30 depicts a line graph showing the effect of 9.2-17 in a B16F10subcutaneous syngeneic model. Tumors were engrafted subcutaneously andtreated with G9.2-17 IgG1 mouse mAb. Animals were dosed on day 0 and day4 intravenously (i.v.) unless otherwise specified in the legend.

FIG. 31 depicts a line graph showing the effect of 9.2-17 in a B16F10subcutaneous syngeneic model. Tumors were engrafted subcutaneously andtreated with G9.2-17 IgG2a mouse mAb. Animals were dosed on day 0 andonce every 4 days thereafter until the end of the experiment. mAbs wereadministered i.v. unless otherwise specified in the legend.

FIG. 32 depicts a graph showing a cell based binding assay CRL-2134 celllines were incubated with a biotinylated Fab, and bound Fab was detectedusing neutravidin conjugated with DyLight 650. Samples were thenanalyzed using flow cytometry. Strong signals were observed for theGalectin-9 antibody 9.2-17, but not for the isotype controls. The K_(D)(nM) values for the Gal-9 antibodies in the two formats were as follows:G9.2-17 hIgG1: 0.41±0.07; G9.2-17 mIgG1: 2.91±0.66.

FIGS. 33A and 33B depict graphs showing a thermal stabilitydetermination of anti-Galectin-9 antibodies. The first derivative of thefluorescence emission plotted as a function of temperature (−dF/dT). Themelting temperature is represented as the temperature at which a peak isobserved. Thermal transition was determined using change in binding offluorophor SYPRO Orange (ThermoFisher) using a real-time PCR instrumentwith a heating rate of 1° C. per minute, essentially following a methodas described in Vedadi et al., Chemical screening methods to identifyligands that promote protein stability, protein crystallization, andstructure determination; Proc Natl Acad Sci USA. 2006 Oct. 24;103(43):15835-40.

DETAILED DESCRIPTION OF INVENTION

Provided herein are antibodies capable of binding to Galectin-9 (e.g.,human, mouse, or both). In some embodiments, the anti-Galectin-9antibodies bind to one or more epitopes in the CRD1 and/or CRD2 domains.Such anti-Galectin-9 antibodies are capable of suppressing the signalingmediated by Galectin-9 (e.g., the signaling pathway mediated byGalectin-9/Dectin-1 or Galectin-9/Tim-3) or eliminating pathologic cellsexpressing Galectin-9 via, e.g., ADCC. Accordingly, the anti-Galectin-9antibodies described herein can be used for inhibiting any of theGalectin-9 signaling and/or eliminating Galectin-9 positive pathologiccells, thereby benefiting treatment of diseases associated withGalectin-9, for example, autoimmune diseases, solid tumors, allergicdisorders, or hematological disorders such as hematologicalmalignancies.

Galectin-9, a tandem-repeat lectin, is a beta-galactoside-bindingprotein, which has been shown to have a role in modulating cell-cell andcell-matrix interactions. It is found to be strongly overexpressed inHodgkin's disease tissue and in other pathologic states. It may also befound circulating in the tumor microenvironment (TME).

Galectin-9 is found to interact with Dectin-1, an innate immune receptorwhich is highly expressed on macrophages in PDA, as well as on cancercells (Daley D, et al. Dectin 1 activation on macrophages by galectin 9promotes pancreatic carcinoma and peritumoral immune tolerance; Nat Med.2017; 23(5):556-6). Regardless of the source of Galectin-9, disruptionof its interaction with Dectin-1 has been shown to lead to thereprogramming of CD4⁺ and CD8⁺ cells into indispensable mediators ofanti-tumor immunity. Thus, Galectin-9 serves as a valuable therapeutictarget for blocking the signaling mediated by Dectin-1. Accordingly, insome embodiments, the anti-Galectin-9 antibodies describe herein disruptthe interaction between Galectin-9 and Dectin-1.

Galectin-9 is also found to interact with TIM-3, a type I cell surfaceglycoprotein expressed on the surface of leukemic stem cells in allvarieties of acute myeloid leukemia (except for M3 (acute promyelocyticleukemia)), but not expressed in normal human hematopoietic stem cells(HSCs). TIM-3 signaling resulting from Galectin-9 ligation has beenfound to have a pleiotropic effect on immune cells, inducing apoptosisin Th1 cells (Zhu et al., Nat Immunol., 2005, 6:1245-1252) andstimulating the secretion of tumor necrosis factor-α (TNF-α), leading tothe maturation of monocytes into dendritic cells, resulting ininflammation by innate immunity (Kuchroo et al., Nat Rev Immunol., 2008,8:577-580). Further Galectin-9/TIM-3 signaling has been found toco-activate NF-κB and β-catenin signaling, two pathways that promote LSCself-renewal (Kikushige et al., Cell Stem Cell, 2015, 17(3):341-352). Ananti-Galectin-9 antibody that interferes with Galectin-9/TIM-3 bindingcould have a therapeutic effect, especially with respect to leukemia andother hematological malignancies. Accordingly, in some embodiments, theanti-Galectin-9 antibodies described herein disrupt the interactionbetween Galectin-9 and TIM-3.

Galectin-9 is also found to interact with CD206, a mannose receptorhighly expressed on M2 polarized macrophages, thereby promoting tumorsurvival (Enninga et al., CD206-positive myeloid cells bind galectin-9and promote a tumor-supportive microenvironment. J Pathol. 2018 August;245(4):468-477). Tumor-associated macrophages expressing CD206 aremediators of tumor immunosuppression, angiogenesis, metastasis, andrelapse (see, e.g., Scodeller et al., Precision Targeting of TumorMacrophages with a CD206 Binding Peptide. M1 and M2 had been describedas the functional states of macrophages; Sci Rep. 2017 Nov. 7;7(1):14655, and references therein). Specifically, M1 (also termedclassically activated macrophages) are trigged by Th1-related cytokinesand bacterial products, express high levels of IL-12, and aretumoricidal. By contrast, M2 (so-called alternatively activatedmacrophages) are activated by Th2-related factors, express high level ofanti-inflammatory cytokines, such as IL-10, and facilitate tumorprogression (Biswas and Mantovani; Macrophage plasticity and interactionwith lymphocyte subsets: cancer as a paradigm; Nat Immunol. 2010October; 11(10):889-96). The pro-tumoral effects of M2 include thepromotion of angiogenesis, advancement of invasion and metastasis, andthe protection of the tumor cells from chemotherapy-induced apoptosis(Hu et al., Functional significance of macrophages in pancreatic cancerbiology; Tumour Biol. 2015 December; 36(12): 9119-9126, and referencestherein). Tumor-associated macrophages are thought be of M2-likephenotype and have a protumor role. Galectin-9 has been shown to mediatemyeloid cell differentiation toward an M2 phenotype (Enninga et al.,Galectin-9 modulates immunity by promoting Th2/M2 differentiation andimpacts survival in patients with metastatic melanoma; Melanoma Res.2016 October; 26(5):429-41). It is possible that Galectin-9 bindingCD206 may result in reprogramming TAMs towards the M2 phenotype, similarto what has been previously shown for Dectin. Without wishing to bebound by theory, blocking the interaction of Galectin-9 with CD206 mayprovide one mechanism by which an anti-Galectin antibody, e.g., esdescribed herein in Table 1 and Table 2, such as antibody 9.1-8m13and/or antibody 9.2-17, can be therapeutically beneficial. Accordingly,in some embodiments, the anti-Galectin-9 antibodies described hereindisrupt the interaction between Galectin-9 and CD206.

Galectin-9 has also been shown to interact with protein disulfideisomerase (PDI) and 4-1BB (Bi S, et al. Galectin-9 binding to cellsurface protein disulfide isomerase regulates the redox environment toenhance T-cell migration and HIV entry; Proc Natl Acad Sci USA. 2011;108(26):10650-5; Madireddi et al. Galectin-9 controls the therapeuticactivity of 4-1BB-targeting antibodies. J Exp Med. 2014;211(7):1433-48).

Anti-Galectin-9 antibodies can serve as therapeutic agents for treatingdiseases associated with Galectin-9 (e.g., those in which a Galectin-9signaling plays a role). Without being bound by theory, ananti-Galectin-9 antibody may block a signaling pathway mediated byGalectin-9. For example, the antibody may interfere with the interactionbetween Galectin-9 and its binding partner (e.g., Dectin-1, TIM-3 orCD206), thereby blocking the signaling triggered by theGalectin-9/Ligand interaction. Alternatively, or in addition, ananti-Galectin-9 antibody may also exert its therapeutic effect byinducing blockade and/or cytotoxicity, for example, ADCC, CDC, or ADCPagainst pathologic cells that express Galectin-9. A pathologic cellrefers to a cell that contributes to the initiation and/or developmentof a disease, either directly or indirectly.

Accordingly, described herein are anti-Galectin-9 antibodies andtherapeutic uses thereof for treating diseases associated withGalectin-9.

Antibodies Binding to Galectin-9

The present disclosure provides antibodies that bind Galectin-9, forexample, human and/or mouse Galectin-9.

In some instances, the anti-Galectin antibody described herein binds toan epitope in a carbohydrate recognition domain (CRD) of Galectin-9,e.g., CRD1 or CRD2. In some instances, the anti-Galectin antibody maybind to CRD1 and CRD2. Galectin-9 is a protein well known in the art.For example, NCBI GenBank Accession Nos. BAB83625.1 and NP_034838.2provide information for human and mouse Galectin-1, respectively.Provided herein are exemplary human and mouse Galectin-9 polypeptides;Human galectin-9 (isoform 1; aka “long;”) is provided as SEQ ID NO: 1;human CRD1 and CRD2 are provided herein as SEQ ID NO: 3 and SEQ ID NO:4, respectively; mouse galectin-9 (isoform 1; aka “long;”) is providedas SEQ ID NO: 2; human and mouse CRD1 and CRD2 are provided herein asSEQ ID NO: 5 and SEQ ID NO: 6, respectively.

The CRD1 domain of human Galectin-9 (SEQ ID NO: 3) encompasses residues1-148 of SEQ ID NO:1, and the CRD2 domain (SEQ ID NO: 4) spans residues218-355 of SEQ ID NO: 1. Similarly, the CRD1 domain of murine Galectin-9(SEQ ID NO: 5) spans residues 1-147 of SEQ ID NO:2, and the CRD2 domain(SEQ ID NO: 6) spans residues 226-353 of SEQ ID NO: 2.

Galectin-9 polypeptides from other species are known in the art and canbe obtained from publicly available gene database, for example, GenBank,using either the human sequence or the mouse sequence as a query. TheCRD1 and CRD2 domains of a Galectin-9 polypeptide can be identified byaligning the sequence of that Galectin-9 polypeptide with that of thehuman or mouse Galectin-9 as described herein.

The antibodies described herein bind Galectin-9 or a fragment thereof(e.g., CRD1 or CRD2). As used herein, the term “anti-Galectin-9antibody” refers to any antibody capable of binding to a Galectin-9polypeptide, which can be of a suitable source, for example, human or anon-human mammal (e.g., mouse, rat, rabbit, primate such as monkey,etc.). In some embodiments, the anti-Galectin-9 antibody can be usedtherapeutically to suppress the bioactivity of Galectin-9. In someembodiments, the anti-Galectin-9 antibody may be used in research or maybe used in diagnostic/prognostic methods, e.g., for the detection ofcells expressing Galectin-9 in an assessment of treatment eligibilityand/or efficacy. Alternatively, or in addition, an anti-Galectin-9antibody may block the interaction between Galactin-9 and its ligand(e.g., Dectin-1, TIM-3), thereby suppressing the signaling pathwaytriggered by, for example, the Galactin-9/Dectin-1 or Galectin-9/TIM-3interaction. An anti-Galectin-9 antibody may also elicit the death ofcells expressing Galectin-9, for example, through an antibody-dependentcellular cytotoxicity (ADCC) mechanism.

An antibody (interchangeably used in plural form) is an immunoglobulinmolecule capable of specific binding to a target, such as acarbohydrate, polynucleotide, lipid, polypeptide, etc., through at leastone antigen recognition site, located in the variable region of theimmunoglobulin molecule. As used herein, the term “antibody”, e.g.,anti-Galectin-9 antibody, encompasses not only intact (e.g.,full-length) polyclonal or monoclonal antibodies, but alsoantigen-binding fragments thereof (such as Fab, Fab′, F(ab′)2, Fv),single chain (scFv), mutants thereof, fusion proteins comprising anantibody portion, humanized antibodies, chimeric antibodies, diabodies,nanobodies, linear antibodies, single chain antibodies, multispecificantibodies (e.g., bispecific antibodies) and any other modifiedconfiguration of the immunoglobulin molecule that comprises an antigenrecognition site of the required specificity, including glycosylationvariants of antibodies, amino acid sequence variants of antibodies, andcovalently modified antibodies. An antibody, e.g., anti-Galectin-9antibody, includes an antibody of any class, such as IgD, IgE, IgG, IgA,or IgM (or sub-class thereof), and the antibody need not be of anyparticular class. Depending on the antibody amino acid sequence of theconstant domain of its heavy chains, immunoglobulins can be assigned todifferent classes. There are five major classes of immunoglobulins: IgA,IgD, IgE, IgG, and IgM, and several of these may be further divided intosubclasses (isotypes), e.g., IgG1, IgG2, IgG3, IgG4, IgA1 and IgA2. Theheavy-chain constant domains that correspond to the different classes ofimmunoglobulins are called alpha, delta, epsilon, gamma, and mu,respectively. The subunit structures and three-dimensionalconfigurations of different classes of immunoglobulins are well known.

A typical antibody molecule comprises a heavy chain variable region(V_(H)) and a light chain variable region (V_(L)), which are usuallyinvolved in antigen binding. The V_(H) and V_(L) regions can be furthersubdivided into regions of hypervariability, also known as“complementarity determining regions” (“CDR”), interspersed with regionsthat are more conserved, which are known as “framework regions” (“FR”).Each V_(H) and V_(L) is typically composed of three CDRs and four FRs,arranged from amino-terminus to carboxy-terminus in the following order:FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4. The extent of the framework regionand CDRs can be precisely identified using methodology known in the art,for example, by the Kabat definition, the Chothia definition, the AbMdefinition, and/or the contact definition, all of which are well knownin the art. See, e.g., Kabat, E. A., et al. (1991) Sequences of Proteinsof Immunological Interest, Fifth Edition, U.S. Department of Health andHuman Services, NIH Publication No. 91-3242, Chothia et al., (1989)Nature 342:877; Chothia, C. et al. (1987) J. Mol. Biol. 196:901-917,Al-lazikani et al (1997) J. Molec. Biol. 273:927-948; and Almagro, J.Mol. Recognit. 17:132-143 (2004). See also hgmp.mrc.ac.uk andbioinf.org.uk/abs).

The anti-Galectin-9 antibody described herein may be a full-lengthantibody, which contains two heavy chains and two light chains, eachincluding a variable domain and a constant domain. Alternatively, theanti-Galectin-9 antibody can be an antigen-binding fragment of afull-length antibody. Examples of binding fragments encompassed withinthe term “antigen-binding fragment” of a full length antibody include(i) a Fab fragment, a monovalent fragment consisting of the V_(L),V_(H), C_(L) and C_(H)1 domains; (ii) a F(ab′)₂ fragment, a bivalentfragment including two Fab fragments linked by a disulfide bridge at thehinge region; (iii) a Fd fragment consisting of the V_(H) and C_(H)1domains; (iv) a Fv fragment consisting of the V_(L) and V_(H) domains ofa single arm of an antibody, (v) a dAb fragment (Ward et al., (1989)Nature 341:544-546), which consists of a V_(H) domain; and (vi) anisolated complementarity determining region (CDR) that retainsfunctionality. Furthermore, although the two domains of the Fv fragment,V_(L) and V_(H), are coded for by separate genes, they can be joined,using recombinant methods, by a synthetic linker that enables them to bemade as a single protein chain in which the V_(L) and V_(H) regions pairto form monovalent molecules known as single chain Fv (scFv). See e.g.,Bird et al. (1988) Science 242:423-426; and Huston et al. (1988) Proc.Natl. Acad. Sci. USA 85:5879-5883.

The anti-Galectin-9 antibody as described herein, e.g., in Table 1and/or Table 2, can bind and inhibit (e.g., reduce or eliminate) theactivity of Galectin-9. In some embodiments, the anti-Galectin-9antibody as described herein can bind and inhibit the activity ofGalectin-9 by at least 30% (e.g., 31%, 35%, 40%, 45%, 50%, 60%, 70%,80%, 90%, 95% or greater, including any increment therein). The apparentinhibition constant (Ki^(app) or K_(i,app)), which provides a measure ofinhibitor potency, is related to the concentration of inhibitor requiredto reduce enzyme activity and is not dependent on enzyme concentrations.The inhibitory activity of an anti-Galectin-9 antibody described hereincan be determined by routine methods known in the art.

The K_(i,) ^(app) value of an antibody may be determined by measuringthe inhibitory effect of different concentrations of the antibody on theextent of the reaction (e.g., enzyme activity); fitting the change inpseudo-first order rate constant (v) as a function of inhibitorconcentration to the modified Morrison equation (Equation 1) yields anestimate of the apparent Ki value. For a competitive inhibitor, theKi^(app) can be obtained from the y-intercept extracted from a linearregression analysis of a plot of K_(i,) ^(app) versus substrateconcentration.

$\begin{matrix}{v = {A \cdot \frac{\begin{matrix}{\left( {\lbrack E\rbrack - \lbrack I\rbrack - K_{i}^{app}} \right) +} \\{\sqrt{\left( {\lbrack E\rbrack - \lbrack I\rbrack - K_{i}^{app}} \right)^{2} + {{4\lbrack E\rbrack} \cdot}}K_{i}^{app}}\end{matrix}}{2}}} & \left( {{Equation}\mspace{14mu} 1} \right)\end{matrix}$

Where A is equivalent to v_(o)/E, the initial velocity (v_(o)) of theenzymatic reaction in the absence of inhibitor (I) divided by the totalenzyme concentration (E). In some embodiments, the anti-Galectin-9antibody described herein may have a Ki^(app) value of 1000, 900, 800,700, 600, 500, 400, 300, 200, 100, 50, 40, 30, 20, 19, 18, 17, 16, 15,14, 13, 12, 11, 10, 9, 8, 7, 6, 5 pM or less for the target antigen orantigen epitope. In some embodiments, the anti-Galectin-9 antibody mayhave a lower Ki^(app) for a first target (e.g., the CRD2 of Galectin-9)relative to a second target (e.g., CRD1 of Galectin-9). Differences inKi^(app) (e.g., for specificity or other comparisons) can be at least1.5, 2, 3, 4, 5, 10, 15, 20, 37.5, 50, 70, 80, 91, 100, 500, 1000,10,000 or 10⁵ fold. In some examples, the anti-Galectin-9 antibodyinhibits a first antigen (e.g., a first protein in a first conformationor mimic thereof) greater relative to a second antigen (e.g., the samefirst protein in a second conformation or mimic thereof; or a secondprotein). In some embodiments, any of the anti-Galectin-9 antibodies maybe further affinity matured to reduce the Ki^(app) of the antibody tothe target antigen or antigenic epitope thereof.

In some embodiments, the anti-Galectin-9 antibody suppresses theDectin-1 signaling, e.g., in tumor infiltrating immune cells, such asmacrophages. In some embodiments, the anti-Galectin-9 antibodysuppresses the Dectin-1 signaling triggered by Galectin-9 by at least30% (e.g., 31%, 35%, 40%, 50%, 60%, 70%, 80%, 90%, 95% or greater,including any increment therein). Such inhibitory activity can bedetermined by conventional methods or the assays described herein, forexample, Example 2. Alternatively or in addition, the anti-Galectin-9antibody may suppress the T cell immunoglobulin mucin-3 (TIM-3)signaling initiated by Galectin-9. In some embodiments, theanti-Galectin-9 antibody suppresses the T cell immunoglobulin mucin-3(TIM-3) signaling, e.g., in tumor infiltrating immune cells, e.g., insome embodiments by at least 30% (e.g., 31%, 35%, 40%, 50%, 60%, 70%,80%, 90%, 95% or greater, including any increment therein). Suchinhibitory activity can be determined by conventional methods or theassays described herein, for example, Example 2.

In some embodiments, the anti-Galectin-9 antibody suppresses the CD206signaling, e.g., in tumor infiltrating immune cells. In someembodiments, the anti-Galectin-9 antibody suppresses the CD206 signalingtriggered by Galectin-9 by at least 30% (e.g., 31%, 35%, 40%, 50%, 60%,70%, 80%, 90%, 95% or greater, including any increment therein). Suchinhibitory activity can be determined by conventional methods or theassays described herein, for example, Example 13. In some embodiments,the anti-Galectin-9 antibody blocks or prevents binding of Galectin-9 toCD206 by at least 30% (e.g., 31%, 35%, 40%, 50%, 60%, 70%, 80%, 90%, 95%or greater, including any increment therein). Such inhibitory activitycan be determined by conventional methods or the assays describedherein, for example, Example 13.

In some embodiments, any of the anti-Galectin-9 antibodies describedherein induce cell cytotoxicity, such as ADCC, in target cellsexpressing Galectin-9, e.g., wherein the target cells are cancer cellsor immune suppressive immune cells. In some embodiments, theanti-Galectin-9 antibody induces apoptosis in immune cells, such as Tcells, or cancer cells by at least 30% (e.g., 31%, 35%, 40%, 50%, 60%,70%, 80%, 90%, 95% or greater, including any increment therein). Suchinhibitory activity can be determined by conventional methods or theassays described herein, for example, Example 14. In some embodiments,any of the anti-Galectin-9 antibodies described herein induce cellcytotoxicity such as complement-dependent cytotoxicity (CDC) againsttarget cells expressing Galectin-9.

Antibody-dependent cell-mediated phagocytosis (ADCP) is an importantmechanism of action for antibodies that mediate part or all of theiraction though phagocytosis. In that case, antibodies mediate uptake ofspecific antigens by antigen presenting cells. ADCP can be mediated bymonocytes, macrophages, neutrophils, and dendritic cells, throughFcγRIIa, FcγRI, and FcγRIIIa, of which FcγRIIa (CD32a) on macrophagesrepresent the predominant pathway. In some embodiments, any of theanti-Galectin-9 antibodies described herein induce cell phagocytosis oftarget cells, e.g., cancer cells or immune suppressive immune cellsexpressing Galectin-9 (ADCP). In some embodiments, the anti-Galectin-9antibody increases phagocytosis of target cells, e.g., cancer cells orimmune suppressive immune cells, by at least 30% (e.g., 31%, 35%, 40%,50%, 60%, 70%, 80%, 90%, 95% or greater, including any incrementtherein).

In some embodiments, any of the anti-Galectin-9 antibodies describedherein induce cell cytotoxicity such as complement-dependentcytotoxicity (CDC) against target cells, e.g., cancer cells or immunesuppressive immune cells. In some embodiments, the anti-Galectin-9antibody increases CDC against target cells by at least 30% (e.g., 31%,35%, 40%, 50%, 60%, 70%, 80%, 90%, 95% or greater, including anyincrement therein).

In some embodiments, any of the anti-Galectin-9 antibodies describedherein induce T cell activation, e.g., in tumor infiltrating T cells,i.e., suppress Galectin-9 mediated inhibition of T cell activation,either directly or indirectly. In some embodiments, the anti-Galectin-9antibody promotes T cell activation by at least 30% (e.g., 31%, 35%,40%, 50%, 60%, 70%, 80%, 90%, 95% or greater, including any incrementtherein). T cell activation can be determined by conventional methods orthe assays described herein, for example, Example 6 (e.g., measurementof CD44, OX40, IFNgamma, PD-1). In some embodiments, the anti-Galectin-9antibody promotes CD4+ cell activation by at least 30% (e.g., 31%, 35%,40%, 50%, 60%, 70%, 80%, 90%, 95% or greater, including any incrementtherein). In a non-limiting example, the anti-Galectin antibody inducesCD44 expression in CD4+ cells. In some embodiments, the anti-Galectin-9antibody increases CD44 expression in CD4+ cells by at least 30% (e.g.,31%, 35%, 40%, 50%, 60%, 70%, 80%, 90%, 95% or greater, including anyincrement therein). In a non-limiting example, the anti-Galectinantibody induces IFNgamma expression in CD4+ cells. In some embodiments,the anti-Galectin-9 antibody increases IFNgamma expression in CD4+ cellsby at least 30% (e.g., 31%, 35%, 40%, 50%, 60%, 70%, 80%, 90%, 95% orgreater, including any increment therein). In a non-limiting example,the anti-Galectin antibody induces TNFalpha expression in CD4+ cells. Insome embodiments, the anti-Galectin-9 antibody increases TNFalphaexpression in CD4+ cells by at least 30% (e.g., 31%, 35%, 40%, 50%, 60%,70%, 80%, 90%, 95% or greater, including any increment therein).

In some embodiments, the anti-Galectin-9 antibody promotes CD8+ cellactivation by at least 30% (e.g., 31%, 35%, 40%, 50%, 60%, 70%, 80%,90%, 95% or greater), including any increment therein). In anon-limiting example, the anti-Galectin antibody induces CD44 expressionin CD8+ cells. In some embodiments, the anti-Galectin-9 antibodyincreases CD44 expression in CD8+ cells by at least 30% (e.g., 31%, 35%,40%, 50%, 60%, 70%, 80%, 90%, 95% or greater, including any incrementtherein). In a non-limiting example, the anti-Galectin antibody inducesIFNgamma expression in CD8+ cells. In some embodiments, theanti-Galectin-9 antibody increases IFNgamma expression in CD8+ cells byat least 30% (e.g., 31%, 35%, 40%, 50%, 60%, 70%, 80%, 90%, 95% orgreater, including any increment therein). In a non-limiting example,the anti-Galectin antibody induces TNFalpha expression in CD8+ cells. Insome embodiments, the anti-Galectin-9 antibody increases TNFalphaexpression in CD8+ cells by at least 30% (e.g., 31%, 35%, 40%, 50%, 60%,70%, 80%, 90%, 95% or greater, including any increment therein).

The antibodies described herein can be murine, rat, human, or any otherorigin (including chimeric or humanized antibodies). Such antibodies arenon-naturally occurring, i.e., would not be produced in an animalwithout human act (e.g., immunizing such an animal with a desiredantigen or fragment thereof or isolated from antibody libraries).

Any of the antibodies described herein, e.g., anti-Galectin-9 antibody,can be either monoclonal or polyclonal. A “monoclonal antibody” refersto a homogenous antibody population and a “polyclonal antibody” refersto a heterogeneous antibody population. These two terms do not limit thesource of an antibody or the manner in which it is made.

In some embodiments, the anti-Galectin-9 antibody is a humanizedantibody. In some embodiments, the anti-Galectin-9 antibody is ahumanized antibody having one of more of the elements or characteristicsdescribed below or elsewhere herein. Humanized antibodies refer to formsof non-human (e.g., murine) antibodies that are specific chimericimmunoglobulins, immunoglobulin chains, or antigen-binding fragmentsthereof that contain minimal sequence derived from non-humanimmunoglobulin. In general, humanized antibodies are humanimmunoglobulins (recipient antibody) in which residues from a CDR of therecipient are replaced by residues from a CDR of a non-human species(donor antibody) such as mouse, rat, or rabbit having the desiredspecificity, affinity, and capacity. In some instances, Fv frameworkregion (FR) residues of the human immunoglobulin are replaced bycorresponding non-human residues. Furthermore, the humanized antibodymay comprise residues that are found neither in the recipient antibodynor in the imported CDR or framework sequences, but are included tofurther refine and optimize antibody performance. In some instances, thehumanized antibody may comprise substantially all of at least one, andtypically two, variable domains, in which all or substantially all ofthe CDR regions correspond to those of a non-human immunoglobulin andall or substantially all of the FR regions are those of a humanimmunoglobulin consensus sequence. The humanized antibody optimally alsowill comprise at least a portion of an immunoglobulin constant region ordomain (Fc), typically that of a human immunoglobulin. Antibodies mayhave Fc regions modified as described in WO 99/58572. Other forms ofhumanized antibodies have one or more CDRs (one, two, three, four, five,or six) which are altered with respect to the original antibody, whichare also termed one or more CDRs “derived from” one or more CDRs fromthe original antibody. Humanized antibodies may also involve affinitymaturation.

Methods for constructing humanized antibodies are also well known in theart. See, e.g., Queen et al., Proc. Natl. Acad. Sci. USA, 86:10029-10033(1989). In one example, variable regions of V_(H) and V_(L) of a parentnon-human antibody are subjected to three-dimensional molecular modelinganalysis following methods known in the art. Next, framework amino acidresidues predicted to be important for the formation of the correct CDRstructures are identified using the same molecular modeling analysis. Inparallel, human V_(H) and V_(L) chains having amino acid sequences thatare homologous to those of the parent non-human antibody are identifiedfrom any antibody gene database using the parent V_(H) and V_(L)sequences as search queries. Human V_(H) and V_(L) acceptor genes arethen selected.

The CDR regions within the selected human acceptor genes can be replacedwith the CDR regions from the parent non-human antibody or functionalvariants thereof. When necessary, residues within the framework regionsof the parent chain that are predicted to be important in interactingwith the CDR regions can be used to substitute for the correspondingresidues in the human acceptor genes.

In some embodiments, the anti-Galectin-9 antibody is a chimericantibody. In some embodiments, the anti-Galectin-9 antibody is achimeric antibody which may include a heavy constant region and a lightconstant region from a human antibody. Chimeric antibodies refer toantibodies having a variable region or part of variable region from afirst species and a constant region from a second species. Typically, inthese chimeric antibodies, the variable region of both light and heavychains mimics the variable regions of antibodies derived from onespecies of mammals (e.g., a non-human mammal such as mouse, rabbit, andrat), while the constant portions are homologous to the sequences inantibodies derived from another mammal such as human. In someembodiments, amino acid modifications can be made in the variable regionand/or the constant region.

In some embodiments, the anti-Galectin-9 antibodies described hereinspecifically bind to the corresponding target antigen or an epitopethereof, e.g., Galectin-9 antigen or epitope. An antibody that“specifically binds” to an antigen or an epitope is a term wellunderstood in the art. A molecule is said to exhibit “specific binding”if it reacts more frequently, more rapidly, with greater duration and/orwith greater affinity with a particular target antigen than it does withalternative targets. An antibody “specifically binds” to a targetantigen or epitope if it binds with greater affinity, avidity, morereadily, and/or with greater duration than it binds to other substances.For example, an antibody that specifically (or preferentially) binds toan antigen (Galectin-9) or an antigenic epitope therein is an antibodythat binds this target antigen with greater affinity, avidity, morereadily, and/or with greater duration than it binds to other antigens orother epitopes in the same antigen. It is also understood with thisdefinition that, for example, an antibody that specifically binds to afirst target antigen may or may not specifically or preferentially bindto a second target antigen. As such, “specific binding” or “preferentialbinding” does not necessarily require (although it can include)exclusive binding. In some examples, an antibody that “specificallybinds” to a target antigen or an epitope thereof may not bind to otherantigens or other epitopes in the same antigen (i.e., only baselinebinding activity can be detected in a conventional method). In someembodiments, the anti-Galectin-9 antibodies described hereinspecifically bind to Galectin-9. In some embodiments, theanti-Galectin-9 antibodies described herein specifically bind to theCRD2 of Galectin-9. In some embodiments, the anti-Galectin-9 antibodiesdescribed herein specifically bind to the CRD1 of Galectin-9.Alternatively, or in addition, the anti-Galectin-9 antibody describedherein specifically binds human Galectin-9 or a fragment thereof asrelative to the mouse counterpart, or vice versa (e.g., having a bindingaffinity at least 10-fold higher to one antigen than the other asdetermined in the same assay under the same assay conditions).

In some embodiments, the anti-Galectin-9 antibody binds only to CRD1(and not CRD2), for example, meaningful binding to CRD2 or binding toCRD2 is not detectable by a routine assay method. In some embodiments,the anti-Galectin-9 or a fragment thereof binds only to CRD2 (and notCRD1). In some embodiments, certain antibodies described herein may bindto both CRD1 and CRD2. In some embodiments, certain antibodies orfragments thereof described herein may bind to both CRD1 and CRD2, butwith a lower affinity to CRD2. In some embodiments, certain antibodiesor fragments thereof described herein may bind to both CRD1 and CRD2,but with a lower affinity to CRD1.

In some embodiments, the effect of a CRD1 binding Gal-9 antibody and aCRD2 binding Gal-9 antibody may be additive. In some embodiments, theeffect of a CRD1 binding Gal-9 antibody and a CRD2 binding Gal-9antibody may be synergistic. In some embodiments, a “cocktail” i.e., amixture of two or more antibodies may be used in a composition. Suchcompositions may comprise one or more antibodies that bind to CRD1described herein and one or more antibodies that bind to CRD2 describedherein. In a non-limiting example, an antibody comprising the variableregion of clone 9.1-8m13 (e.g., SEQ ID NO: 21 (light chain and SEQ IDNO: 86) can be combined with an antibody comprising the variable regionof clone 9.2-17 (SEQ ID NO: 54 (light chain and SEQ ID NO: 55) in acomposition. Antibodies may be mixed in equimolar amounts or in otherratios, as determined optimal for performance.

In some embodiments, an antibody might bind to both CRD1 and CRD2. Inother instances, the anti-Galectin-9 antibody described herein maycross-react to human and a non-human Galectin-9 (e.g., mouse), e.g., thedifference in binding affinity to the human and the non-human Galectin-9is less than 5-fold, e.g., less than 2-fold, or substantially similar.

In some embodiments, an anti-Galectin-9 antibody as described herein hasa suitable binding affinity for the target antigen (e.g., Galectin-9) orantigenic epitopes thereof. As used herein, “binding affinity” refers tothe apparent association constant or K_(A). The K_(A) is the reciprocalof the dissociation constant (K_(D)). The anti-Galectin-9 antibodydescribed herein may have a binding affinity (K_(D)) of at least 10⁻⁵,10⁻⁶, 10⁻⁷, 10⁻⁸, 10⁻⁹, 10⁻¹⁰ M, or lower for the target antigen orantigenic epitope. An increased binding affinity corresponds to adecreased K_(D). Higher affinity binding of an antibody for a firstantigen relative to a second antigen can be indicated by a higher K_(A)(or a smaller numerical value K_(D)) for binding the first antigen thanthe K_(A) (or numerical value K_(D)) for binding the second antigen. Insuch cases, the antibody has specificity for the first antigen (e.g., afirst protein in a first conformation or mimic thereof) relative to thesecond antigen (e.g., the same first protein in a second conformation ormimic thereof; or a second protein). In some embodiments, theanti-Galectin-9 antibodies described herein have a higher bindingaffinity (a higher K_(A) or smaller K_(D)) to the CRD1 of Galectin-9 ascompared to the binding affinity to the CRD2 of Galectin-9. In someembodiments, the anti-Galectin-9 antibodies described herein have ahigher binding affinity (a higher K_(A) or smaller K_(D)) to the CRD2 ofGalectin-9 as compared to the binding affinity to the CRD1 ofGalectin-9. Differences in binding affinity (e.g., for specificity orother comparisons) can be at least 1.5, 2, 3, 4, 5, 10, 15, 20, 37.5,50, 70, 80, 91, 100, 500, 1000, 10,000 or 10⁵ fold. In some embodiments,any of the anti-Galectin-9 antibodies may be further affinity matured toincrease the binding affinity of the antibody to the target antigen orantigenic epitope thereof.

Binding affinity (or binding specificity) can be determined by a varietyof methods including equilibrium dialysis, equilibrium binding, gelfiltration, ELISA, surface plasmon resonance, or spectroscopy (e.g.,using a fluorescence assay). Exemplary conditions for evaluating bindingaffinity are in HBS-P buffer (10 mM HEPES pH7.4, 150 mM NaCl, 0.005%(v/v) Surfactant P20).

These techniques can be used to measure the concentration of boundbinding protein as a function of target protein concentration. Undercertain conditions, the fractional concentration of bound bindingprotein ([Bound]/[Total]) is generally related to the concentration oftotal target protein ([Target]) by the following equation:[Bound]/[Total]=[Target]/(Kd+[Target])

It is not always necessary to make an exact determination of K_(A),though, since sometimes it is sufficient to obtain a quantitativemeasurement of affinity, e.g., determined using a method such as ELISAor FACS analysis, is proportional to K_(A), and thus can be used forcomparisons, such as determining whether a higher affinity is, e.g.,2-fold higher, to obtain a qualitative measurement of affinity, or toobtain an inference of affinity, e.g., by activity in a functionalassay, e.g., an in vitro or in vivo assay. In some cases, the in vitrobinding assay is indicative of in vivo activity. In other cases, the invitro binding assay is not necessarily indicative of in vivo activity.In some cases tight binding is beneficial, but in other cases tightbinding may not be as desirable in vivo, and an antibody with lowerbinding affinity may be more desirable. A number of exemplaryanti-Galectin-9 antibodies (specific to CRD1 or CRD2) are providedherein.

Exemplary antibody clones (reference antibodies) of the disclosurebinding to CRD1 include G9.1-1, G9.1-2, G9.1-3, G9.1-4, G9.1-5, G9.1-6,G9.1-7, G9.1-8, G9.1-9, G9.1-10, G9.1-11, G9.1-8m1, G9.1-8m2, G9.1-8m3,G9.1-8m4, G9.1-8m5, G9.1-8m6, G9.1-8m7, G9.1- 8m8, G9.1-8m9, G9.1-8m10,G9.1-8m11, G9.1-8m12, G9.1-8m13, and G9.1-8m14. Exemplary antibodyclones (reference antibodies) of the disclosure binding to CRD2 includeG9.2-1, G9.2-2, G9.2-3, G9.2-4, G9.2-5, G9.2-6, G9.2-7, G9.2-8, G9.2-9,G9.2-10, G9.2-11, G9.2-12, G9.2-13, G9.2-14, G9.2-15, G9.2-16, G9.2-17,G9.2-17mut6, G9.2-18, G9.2-19, G9.2-20, G9.2-21, G9.2-22, G9.2-23,G9.2-24, G9.2-25, G9.2-26, and G9.2-low affinity binder.

Variable Regions

Exemplary anti-Galectin-9 antibodies described herein binding to CRD1are antibodies, e.g., monoclonal, recombinant, and/or human antibodies,having the CDR and/or variable region sequences of antibodies G9.1-1,G9.1-2, G9.1-3, G9.1-4, G9.1-5, G9.1-6, G9.1-7, G9.1-8, G9.1-9, G9.1-10,G9.1-11, G9.1-8m1, G9.1-8m2, G9.1-8m3, G9.1-8m4, G9.1-8m5, G9.1-8m6,G9.1-8m7, G9.1-8m8, G9.1-8m9, G9.1-8m10, G9.1-8m11, G9.1-8m12,G9.1-8m13, and G9.1-8m14. Exemplary anti-Galectin-9 antibodies describedherein binding to CRD2 are antibodies, e.g., monoclonal, recombinant,and/or human antibodies, having the CDR and/or variable region sequencesof antibodies G9.2-1, G9.2-2, G9.2-3, G9.2-4, G9.2-5, G9.2-6, G9.2-7,G9.2-8, G9.2-9, G9.2-10, G9.2-11, G9.2-12, G9.2-13, G9.2-14, G9.2-15,G9.2-16, G9.2-17, G9.2-17mut6, G9.2-18, G9.2-19, G9.2-20, G9.2-21,G9.2-22, G9.2-23, G9.2-24, G9.2-25, G9.2-26, and G9.2-low affinitybinder. Exemplary sequences and SEQ ID NOs are listed in Table 1 and 2.CDRs determined using the Kabat methodology are shown in boldface. Table3 presents the CDRs, determined with Kabat methodology, of selectedclones. Herein the terms “m” and “mut”, e.g., “9.1-8m” and “9.1-8mut”are used interchangeably. For example, the “G9.1-8m1”, “G9.1-8m2”,“G9.1-8m3”, “G9.1-8m4”, “G9.1-8m5”, “G9.1-8m6”, “G9.1-8m7”, “G9.1-8m8”,“G9.1-8m9”, “G9.1-8m10”, “G9.1-8m11”, “G9.1-8m12”, “G9.1-8m13”, and“G9.1-8m14” are used interchangeably with “G9.1-8mut1”, “G9.1-8mut2”,“G9.1-8mut3”, “G9.1-8mut4”, “G9.1-8mut5”, “G9.1-8mut6”, “G9.1-8mut7”,“G9.1-8mut8”, “G9.1-8mut9”, “G9.1-8mut10”, “G9.1-8mut11”, “G9.1-8mut12”,“G9.1-8mut13”, and “G9.1-8mut14, respectively.

TABLE 1 Antibodies directed against CRD1 CDR CDR CDR LC/HC IgG1 IgG4 VR1 2 3 IgG1 LALA IgG4 mut Clone Sequence SEQ ID NO: G9.1-1 V_(L):DIQMTQSPSSLSASVGDRV 7 328 329 330 88 88 88 88 TITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGVPSRF SGSRSGTDFTLTISSLQPEDFAT YYCQQSWVGSLITFGQGTKV EIKRG9.1-1 V_(H): EVQLVESGGGLVQPGGSLR 8 431 438 367 116 169 222 275LSCAASGFTFSSSSIHWVRQAP GKGLEWVASIYSSYGYTYYA DSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARYYW GWSQNQGFWWYGLDYWGQ GTLVTVSS G9.1-2 V_(L):DIQMTQSPSSLSASVGDRV 9 328 329 331 89 89 89 89 TITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGVPSRF SGSRSGTDFTLTISSLQPEDFAT YYCQQWQWGYSLVTFGQGTKVEIKR G9.1-2 V_(H): EVQLVESGGGLVQPGGSLR 10 435 439 368 117 170 223 276LSCAASGFTISSSSIHWVRQAP GKGLEWVASISSYYGSTYYAD SVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARSWSSSF WYNWALDYWGQGTLVTVSS G9.1-3 V_(L):DIQMTQSPSSLSASVGDRV 11 328 329 332 90 90 90 90 TITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGVPSRF SGSRSGTDFTLTISSLQPEDFAT YYCQQSWYSNKPITFGQGTKVEIKR G9.1-3 V_(H): EVQLVESGGGLVQPGGSLR 12 436 363 369 118 171 224 277LSCAASGFTIYSSSIHWVRQAP GKGLEWVAYIYSSSGYTSYAD SVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARYSHSSL YYSWIWALDYWGQGTLVTVS S G9.1-4 V_(L):DIQMTQSPSSLSASVGDRV 13 328 329 333 91 91 91 91 TITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGVPSRF SGSRSGTDFTLTISSLQPEDFAT YYCQQSSSSLITFGQGTKVEIK RG9.1-4 V_(H): EVQLVESGGGLVQPGGSLR 14 437 440 370 119 172 225 278LSCAASGFTIYYSSIHWVRQA PGKGLEWVASISSSSGSTSYA DSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARSYRP YSSYYWGMDYWGQGTLVTV SS G9.1-5 V_(L):DIQMTQSPSSLSASVGDRV 15 328 329 334 92 92 92 92 TITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGVPSRF SGSRSGTDFTLTISSLQPEDFAT YYCQQYYGWFYPVTFGQGTKVEIKR G9.1-5 V_(H): EVQLVESGGGLVQPGGSLR 16 437 441 371 120 173 226 279LSCAASGFTIYYSSIHWVRQA PGKGLEWVASISSSYGSTYYA DSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARSVSW YPYYYYYGYGSGLDYWGQG TLVTVSS G9.1-6 V_(L):DIQMTQSPSSLSASVGDRV 17 328 329 335 93 93 93 93 TITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGVPSRF SGSRSGTDFTLTISSLQPEDFAT YYCQQYHSSLFTFGQGTKVEI KRG9.1-6 V_(H): EVQLVESGGGLVQPGGSLR 18 427 442 372 121 174 227 280LSCAASGFTLSSSSIHWVRQA PGKGLEWVASIYSSYGSTSYA DSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARSSHW YMYWSYWGWYIGMDYWGQ GTLVTVSS G9.1-7 V_(L):DIQMTQSPSSLSASVGDRV 19 328 329 336 94 94 94 94 TITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGVPSRF SGSRSGTDFTLTISSLQPEDFAT YYCQQYPGYRGLITFGQGTKVEIKR G9.1-7 V_(H): EVQLVESGGGLVQPGGSLR 20 361 443 373 122 175 228 281LSCAASGFTVSSSSIHWVRQA PGKGLEWVASISSYYGYTYYA DSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARSYSY GYDYFVKYYTMDYWGQGTL VTVSS G9.1-8 V_(L):DIQMTQSPSSLSASVGDRV 21 328 329 337 95 95 95 95 TITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGVPSRF SGSRSGTDFTLTISSLQPEDFAT YYCQQSYYDSNPITFGQGTKVEIKR G9.1-8 V_(H): EVQLVESGGGLVQPGGSLR 22 361 364 374 123 176 229 282LSCAASGFTVSSSSIHWVRQA PGKGLEWVAYIYPYSGYTSY ADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARYST YSWGGIGKWVWGMDYWGQ GTLVTVSS G9.1-9 V_(L):DIQMTQSPSSLSASVGDRV 23 328 329 338 96 96 96 96 TITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGVPSRF SGSRSGTDFTLTISSLQPEDFAT YYCQQSYFHKIPITFGQGTKVEIKR G9.1-9 V_(H): EVQLVESGGGLVQPGGSLR 24 361 363 384 138 191 244 297LSCAASGFTVSSSSIHWVRQA PGKGLEWVAYIYSSSGYTSYA DSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARYSSY HYPYWLFAMDYWGQGTLVT VSS G9.1-10 V_(L):DIQMTQSPSSLSASVGDRV 25 328 329 339 97 97 97 97 TITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGVPSRF SGSRSGTDFTLTISSLQPEDFAT YYCQQWYWYYPVTFGQGTK VEIKRG9.1-10 V_(H): EVQLVESGGGLVQPGGSLR 26 429 444 385 139 192 245 298LSCAASGFTVSYSSIHWVRQA PGKGLEWVASIYSYYGSTYYA DSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARGHYQ EGRKSGFSYWSPALDYWGQ GTLVTVSS G9.1-11 V_(L):DIQMTQSPSSLSASVGDRV 27 328 329 340 98 98 98 98 TITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGVPSRF SGSRSGTDFTLTISSLQPEDFAT YYCQQTYWGLITFGQGTKVE IKRG9.1-11 V_(H): EVQLVESGGGLVQPGGSLR 28 428 445 386 140 193 246 299LSCAASGFTVYSSSIHWVRQA PGKGLEWVASIYSYYGYTSYA DSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARSTEG YDRWGYYSSYWSSGLDYWG QGTLVTVSS G9.1-8m1 V_(L):DIQMTQSPSSLSASVGDRV 21 328 329 337 95 95 95 95 TITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGVPSRF SGSRSGTDFTLTISSLQPEDFAT YYCQQSYYDSNPITFGQGTKVEIKR G9.1-8m1 V_(H): EVQLVESGGGLVQPGGSLR 74 361 365 374 124 177 230 283LSCAASGFTVSSSSIHWVRQA PGKGLEWVA SSSSSSGYTSYA DSVKG RFTISADTSKNTAYLQMNSLRAEDTAVYYCARYSTY SWGGIGKWVWGMDYWGQG TLVTVSS G9.1-8m2 V_(L):DIQMTQSPSSLSASVGDRV 21 328 329 337 95 95 95 95 TITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGVPSRF SGSRSGTDFTLTISSLQPEDFAT YYCQQSYYDSNPITFGQGTKVEIKR G9.1-8m2 V_(H): EVQLVESGGGLVQPGGSLR 75 361 366 374 125 178 231 284LSCAASGFTVSSSSIHWVRQA PGKGLEWVA YIYPYSSSSSYA DSVKG RFTISADTSKNTAYLQMNSLRAEDTAVYYCARYSTY SWGGIGKWVWGMDYWGQG TLVTVSS G9.1-8m3 V_(L):DIQMTQSPSSLSASVGDRV 21 328 329 337 95 95 95 95 TITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGVPSRF SGSRSGTDFTLTISSLQPEDFAT YYCQQSYYDSNPITFGQGTKVEIKR G9.1-8m3 V_(H): EVQLVESGGGLVQPGGSLR 76 361 364 375 126 179 232 285LSCAASGFTVSSSSIHWVRQA PGKGLEWVAYIYPYSGYTSY ADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCAR SSSS SWGGIGKWVWGMDY WGQG TLVTVSS G9.1-8m4 V_(L):DIQMTQSPSSLSASVGDRV 21 328 329 337 95 95 95 95 TITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGVPSRF SGSRSGTDFTLTISSLQPEDFAT YYCQQSYYDSNPITFGQGTKVEIKR G9.1-8m4 V_(H): EVQLVESGGGLVQPGGSLR 77 361 364 376 127 180 233 286LSCAASGFTVSSSSIHWVRQA PGKGLEWVAYIYPYSGYTSY ADSVKGRFTI SADTSKNTAYLQMNSLRAEDTAVYYCAR YST YSSSSSSKWVWGMDYWGQG TLVTVSS G9.1-8m5 V_(L):DIQMTQSPSSLSASVGDRV 21 328 329 337 95 95 95 95 TITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGVPSRF SGSRSGTDFTLTISSLQPEDFAT YYCQQSYYDSNPITFGQGTKVEIKR G9.1-8m5 V_(H): EVQLVESGGGLVQPGGSLR 78 361 364 377 128 181 234 287LSCAASGFTVSSSSIHWVRQA PGKGLEWVAYIYPYSGYTSY ADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCAR YST YSWGGIGSSSSSMDY WGQGT LVTVSS G9.1-8m6 V_(L):DIQMTQSPSSLSASVGDRV 21 328 329 337 95 95 95 95 TITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGVPSRF SGSRSGTDFTLTISSLQPEDFAT YYCQQSYYDSNPITFGQGTKVEIKR G9.1-8m6 V_(H): EVQLVESGGGLVQPGGSLR 79 361 364 378 129 182 235 288LSCAASGFTVSSSSIHWVRQA PGKGLEWVAYIYPYSGYTSY ADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCAR YST YSSSSSKWVWGMDY WGQGT LVTVSS G9.1-8m7 V_(L):DIQMTQSPSSLSASVGDRV 21 328 329 337 95 95 95 95 TITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGVPSRF SGSRSGTDFTLTISSLQPEDFAT YYCQQSYYDSNPITFGQGTKVEIKR G9.1-8m7 V_(H): EVQLVESGGGLVQPGGSLR 80 361 364 379 130 183 236 289LSCAASGFTVSSSSIHWVRQA PGKGLEWVAYIYPYSGYTSY ADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCAR YST YSSSSKWVWGMDY WGQGTL VTVSS G9.1-8m8 V_(L):DIQMTQSPSSLSASVGDRV 21 328 329 f337 95 95 95 95 TITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGVPSRF SGSRSGTDFTLTISSLQPEDFAT YYCQQSYYDSNPITFGQGTKVEIKR G9.1-8m8 V_(H): EVQLVESGGGLVQPGGSLR 81 361 364 380 131 184 237 290LSCAASGFTVSSSSIHWVRQA PGKGLEWVAYIYPYSGYTSY ADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCAR YST YSSSKWVWGMDY WGQGTLV TVSS G9.1-8m9 V_(L):DIQMTQSPSSLSASVGDRV 21 328 329 337 95 95 95 95 TITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGVPSRF SGSRSGTDFTLTISSLQPEDFAT YYCQQSYYDSNPITFGQGTKVEIKR G9.1-8m9 V_(H): EVQLVESGGGLVQPGGSLR 82 361 364 383 132 185 238 291LSCAASGFTVSSSSIHWVRQA PGKGLEWVAYIYPYSGYTSY ADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCAR YST YSSKWVWGMDY WGQGTLVT VSS G9.1-8m10 V_(L):DIQMTQSPSSLSASVGDRV 21 328 329 337 95 95 95 95 TITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGVPSRF SGSRSGTDFTLTISSLQPEDFAT YYCQQSYYDSNPITFGQGTKVEIKR G9.1-8m10 V_(H): EVQLVESGGGLVQPGGSLR 83 361 364 381 133 186 239 292LSCAASGFTVSSSSIHWVRQA PGKGLEWVAYIYPYSGYTSY ADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCAR YST YSKWVWGMDY WGQGTLVT VSS G9.1-8m11 V_(L):DIQMTQSPSSLSASVGDRV 21 328 329 337 95 95 95 95 TITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGVPSRF SGSRSGTDFTLTISSLQPEDFAT YYCQQSYYDSNPITFGQGTKVEIKR G9.1-8m11 V_(H): EVQLVESGGGLVQPGGSLR 84 361 364 382 134 187 240 293LSCAASGFTVSSSSIHWVRQA PGKGLEWVAYIYPYSGYTSY ADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCAR YST YKWVWGMDY WGQGTLVTV SS G9.1-8m12 V_(L):DIQMTQSPSSLSASVGDRV 21 328 329 337 95 95 95 95 TITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGVPSRF SGSRSGTDFTLTISSLQPEDFAT YYCQQSYYDSNPITFGQGTKVEIKR G9.1-8m12 V_(H): EVQLVESGGGLVQPGGSLR 85 361 366 380 135 188 241 294LSCAASGFTVSSSSIHWVRQA PGKGLEWVA YIYPYSSSSSYA DSVKG RFTISADTSKNTAYLQMNSLRAEDTAVYYCAR YSTY SSSKWVWGMDY WGQGTLVT VSS G9.1-8m13 V_(L):DIQMTQSPSSLSASVGDRV 21 328 329 337 95 95 95 95 TITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGVPSRF SGSRSGTDFTLTISSLQPEDFAT YYCQQSYYDSNPITFGQGTKVEIKR G9.1-8m13 V_(H): EVQLVESGGGLVQPGGSLR 86 361 366 383 136 189 242 295LSCAASGFTVSSSSIHWVRQA PGKGLEWVA YIYPYSSSSSYA DSVKG RFTISADTSKNTAYLQMNSLRAEDTAVYYCAR YSTY SSKWVWGMDY WGQGTLVTV SS G9.1-8m14 V_(L):DIQMTQSPSSLSASVGDRV 21 328 329 337 95 95 95 95 TITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGVPSRF SGSRSGTDFTLTISSLQPEDFAT YYCQQSYYDSNPITFGQGTKVEIKR G9.1-8m14 V_(H): EVQLVESGGGLVQPGGSLR 87 361 366 382 137 190 243 296LSCAASGFTVSSSSIHWVRQA PGKGLEWVA YIYPYSSSSSYA DSVKG RFTISADTSKNTAYLQMNSLRAEDTAVYYCARYSTY KWVWGMDY WGQGTLVTVSS

TABLE 2 Antibodies directed against CRD2 CDR CDR CDR LC/HC IgG1 IgG4 VR1 2 3 IgG1 LALA IgG4 mut Clone Sequence SEQ ID NO: G9.2-1 V_(L):DIQMTQSPSSLSASVGDRVTI 29 328 329 341 99 99 99 99 TCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGVPSRFSGS RSGTDFTLTISSLQPEDFATYYC QQYKSKYPFTFGQGTKVEIKRG9.2-1 V_(H): EVQLVESGGGLVQPGGSLR 30 424 446 390 141 194 247 300LSCAASGFTLYSSSIHWVRQAP GKGLEWVASIYSSSGYTYYAD SVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARTYTWKSS WSYQTGYGLDYWGQGTLVTV SS G9.2-2 V_(L):DIQMTQSPSSLSASVGDRVTI 13 328 329 333 91 91 91 91 TCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGVPSRFSGS RSGTDFTLTISSLQPEDFATYYC QQSSSSLITFGQGTKVEIKRG9.2-2 V_(H): EVQLVESGGGLVQPGGSLR 31 431 447 391 142 195 248 301LSCAASGFTFSSSSIHWVRQAP GKGLEWVASISPYYGSTYYAD SVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARAVYYYVY NRSWYWWSGGFDYWGQGTL VTVSS G9.2-3 V_(L):DIQMTQSPSSLSASVGDRVTI 13 328 329 333 91 91 91 91 TCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGVPSRFSGS RSGTDFTLTISSLQPEDFATYYC QQSSSSLITFGQGTKVEIKRG9.2-3 V_(H): EVQLVESGGGLVQPGGSLR 32 431 448 392 143 196 249 302LSCAASGFTFSSSSIHWVRQAP GKGLEWVASISSSSGSTSYADS VKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARPAYSYPYY YFHYGAMDYWGQGTLVTVSS G9.2-4 V_(L):DIQMTQSPSSLSASVGDRVTI 13 328 329 342 91 91 91 91 TCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGVPSRFSGS RSGTDFTLTISSLQPEDFATYYC QQSSSSLITFGQGTKVEIKRG9.2-4 V_(H): EVQLVESGGGLVQPGGSLR 33 431 449 393 144 197 250 303LSCAASGFTFSSSSIHWVRQAP GKGLEWVASIYPSYGYTSYAD SVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARAWYHHE YWGHYSGMDYWGQGTLVTVS S G9.2-5 V_(L):DIQMTQSPSSLSASVGDRVTI 34 328 329 343 100 100 100 100TCRASQSVSSAVAWYQQKPGK APKLLIYSASSLYSGVPSRFSGS RSGTDFTLTISSLQPEDFATYYCQQSSWGLITFGQGTKVEIKR G9.2-5 V_(H): EVQLVESGGGLVQPGGSLR 35 431 450 394145 198 251 304 LSCAASGFTFSSSSIHWVRQAP GKGLEWVASIYSSYGSTYYADSVKGRFTISADTSKNTAYLQMN SLRAEDTAVYYCARSGYSHPY YSYYSGMDYWGQGTLVTVSS G9.2-6V_(L): DIQMTQSPSSLSASVGDRVTI 36 328 329 344 101 101 101 101TCRASQSVSSAVAWYQQKPGK APKLLIYSASSLYSGVPSRFSGS RSGTDFTLTISSLQPEDFATYYCQQFWGSKLFTFGQGTKVEIKR G9.2-6 V_(H): EVQLVESGGGLVQPGGSLR 37 431 451 395146 199 252 305 LSCAASGFTFSSSSIHWVRQAP GKGLEWVASIYSYSGYTYYADSVKGRFTISADTSKNTAYLQMN SLRAEDTAVYYCARTYMAGY KYYFISGYGFDYWGQGTLVTV SSG9.2-7 V_(L): DIQMTQSPSSLSASVGDRVTI 38 328 329 345 102 102 102 102TCRASQSVSSAVAWYQQKPGK APKLLIYSASSLYSGVPSRFSGS RSGTDFTLTISSLQPEDFATYYCQQMYYPGYLITFGQGTKVEIK R G9.2-7 V_(H): EVQLVESGGGLVQPGGSLR 39 425 452 396147 200 253 306 LSCAASGFTFSYSSIHWVRQAP GKGLEWVASIYPSYGYTYYADSVKGRFTISADTSKNTAYLQMN SLRAEDTAVYYCARYWDYGW MYFDPAMDYWGQGTLVTVSS G9.2-8V_(L): DIQMTQSPSSLSASVGDRVTI 40 328 329 346 103 103 103 103TCRASQSVSSAVAWYQQKPGK APKLLIYSASSLYSGVPSRFSGS RSGTDFTLTISSLQPEDFATYYCQQDRWWSALTFGQGTKVEIK R G9.2-8 V_(H): EVQLVESGGGLVQPGGSLR 41 425 453 397148 201 254 307 LSCAASGFTFSYSSIHWVRQAP GKGLEWVASIYSYSGYTSYADSVKGRFTISADTSKNTAYLQMN SLRAEDTAVYYCARYMENWE WPYHSAMDYWGQGTLVTVSS G9.2-9V_(L): DIQMTQSPSSLSASVGDRVTI 42 328 329 347 104 104 104 104TCRASQSVSSAVAWYQQKPGK APKLLIYSASSLYSGVPSRFSGS RSGTDFTLTISSLQPEDFATYYCQQSYGSWYPITFGQGTKVEIKR G9.2-9 V_(H): EVQLVESGGGLVQPGGSLR 43 426 454 398149 202 255 308 LSCAASGFTFYSSSIHWVRQAP GKGLEWVASIYSSYGSTYYADSVKGRFTISADTSKNTAYLQMN SLRAEDTAVYYCARSWWYPY WQYYPGGWHSSGFDYWGQG TLVTVSSG9.2-10 V_(L): DIQMTQSPSSLSASVGDRVTI 44 328 329 348 105 105 105 105TCRASQSVSSAVAWYQQKPGK APKLLIYSASSLYSGVPSRFSGS RSGTDFTLTISSLQPEDFATYYCQQGWYASPITFGQGTKVEIKR G9.2-10 V_(H): EVQLVESGGGLVQPGGSLR 45 426 387 399150 203 256 309 LSCAASGFTFYSSSIHWVRQAP GKGLEWVAYISPSSGYTSYADSVKGRFTISADTSKNTAYLQMNS LRAEDTAVYYCARYTMTYQY YPSGAMDYWGQGTLVTVSS G9.2-11V_(L): DIQMTQSPSSLSASVGDRVTI 46 328 329 349 106 106 106 106TCRASQSVSSAVAWYQQKPGK APKLLIYSASSLYSGVPSRFSGS RSGTDFTLTISSLQPEDFATYYCQQYSSHKYPFTFGQGTKVEIKR G9.2-11 V_(H): EVQLVESGGGLVQPGGSLR 47 432 455 400151 204 257 310 LSCAASGFTIYSSYIHWVRQAP GKGLEWVASIYSSSGYTYYADSVKGRFTISADTSKNTAYLQMN SLRAEDTAVYYCARSYIYYMW QYNYGMSGYGLDYWGQGTLV TVSSG9.2-12 V_(L): DIQMTQSPSSLSASVGDRVTI 48 328 329 350 107 107 107 107TCRASQSVSSAVAWYQQKPGK APKLLIYSASSLYSGVPSRFSGS RSGTDFTLTISSLQPEDFATYYCQQWVYPGSLITFGQGTKVEIK R G9.2-12 V_(H): EVQLVESGGGLVQPGGSLR 49 433 456401 152 205 258 311 LCAASGFTLSYSSIHWVRQAP GKGLEWVASISSSYGYTYYADSVKGRFTISADTSKNTAYLQMN SLRAEDTAVYYCARHSPYYLH SWWWSGLDYWGQGTLVTVSSG9.2-13 V_(L): DIQMTQSPSSLSASVGDRVTI 29 328 329 341 99 99 99 99TCRASQSVSSAVAWYQQKPGK APKLLIYSASSLYSGVPSRFSGS RSGTDFTLTISSLQPEDFATYYCQQYKSKYPFTFGQGTKVEIKR G9.2-13 V_(H): EVQLVESGGGLVQPGGSLR 50 434 362 402153 206 259 312 LSCAASGFTLYYSSIHWVRQAP GKGLEWVASISPSYGSTSYADSVKGRFTISADTSKNTAYLQMNS LRAEDTAVYYCARHSWYYPY YYYALDYWGQGTLVTVSS G9.2-14V_(L): DIQMTQSPSSLSASVGDRVTI 13 328 329 333 91 91 91 91TCRASQSVSSAVAWYQQKPGK APKLLIYSASSLYSGVPSRFSGS RSGTDFTLTISSLQPEDFATYYCQQSSSSLITFGQGTKVEIKR G9.2-14 V_(H): EVQLVESGGGLVQPGGSLR 51 361 457 403154 207 260 313 LSCAASGFTVSSSSIHWVRQAP GKGLEWVASISSSSGYTYYADSVKGRFTISADTSKNTAYLQMNS LRAEDTAVYYCARYWSYPYVY FLAFDYWGQGTLVTVSS G9.2-15V_(L): DIQMTQSPSSLSASVGDRVTI 34 328 329 343 100 100 100 100TCRASQSVSSAVAWYQQKPGK APKLLIYSASSLYSGVPSRFSGS RSGTDFTLTISSLQPEDFATYYCQQSSWGLITFGQGTKVEIKR G9.2-15 V_(H): EVQLVESGGGLVQPGGSLR 52 361 458 404155 208 261 314 LSCAASGFTVSSSSIHWVRQAP GKGLEWVASIYSSSGYTSYADSVKGRFTISADTSKNTAYLQMNS LRAEDTAVYYCARNVENYPY WAWPWGYYGAIDYWGQGTL VTVSSG9.2-16 V_(L): DIQMTQSPSSLSASVGDRVTI 13 328 329 333 91 91 91 91TCRASQSVSSAVAWYQQKPGK APKLLIYSASSLYSGVPSRFSGS RSGTDFTLTISSLQPEDFATYYCQQSSSSLITFGQGTKVEIKR G9.2-16 V_(H): EVQLVESGGGLVQPGGSLR 53 361 459 405156 209 262 315 LSCAASGFTVSSSSIHWVRQAP GKGLEWVASIYSSSGYTYYADSVKGRFTISADTSKNTAYLQMN SLRAEDTAVYYCARTYKWSYY TGYGFDYWGQGTLVTVSS G9.2-17V_(L): DIQMTQSPSSLSASVGDRVTI 54 328 329 352 108 108 108 108TCRASQSVSSAVAWYQQKPGK APKLLIYSASSLYSGVPSRFSGS RSGTDFTLTISSLQPEDFATYYCQQSSTDPITFGQGTKVEIKR G9.2-17 V_(H): EVQLVESGGGLVQPGGSLR 55 361 388 406157 210 263 316 LSCAASGFTVSSSSIHWVRQAP GKGLEWVAYISSSSGYTYYADSVKGRFTISADTSKNTAYLQMN SLRAEDTAVYYCARYWSYPSW WPYRGMDYWGQGTLVTVSSG9.2-17mut6 V_(L): DIQMTQSPSSLSASVGDRVTI 54 328 329 352 108 108 108 108TCRASQSVSSAVAWYQQKPGK APKLLIYSASSLYSGVPSRFSGS RSGTDFTLTISSLQPEDFATYYCQQSSTDPITFGQGTKVEIKR G9.2-17mut6 V_(H): EVQLVESGGGLVQPGGSLR 56 361 388407 158 211 264 317 (mutation is LSCAASGFTVSSSSIHWVRQAP underlined)GKGLEWVAYISSSSGYTYYAD SVKGRFTISADTSKNTAYLQMN SLRAEDTAVYYCARYWSYPSW SPYRGMDYWGQGTLVTVSS G9.2-18 V_(L): DIQMTQSPSSLSASVGDRVTI 13 328 329 33391 91 91 91 TCRASQSVSSAVAWYQQKPGK APKLLIYSASSLYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYC QQSSSSLITFGQGTKVEIKR G9.2-18 V_(H):EVQLVESGGGLVQPGGSLR 57 430 363 408 159 212 265 318LSCAASGFTVSSYSIHWVRQAP GKGLEWVAYIYSSSGYTSYAD SVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARVGYYYPY LYLGDGLDYWGQGTLVTVSS G9.2-19 V_(L):DIQMTQSPSSLSASVGDRVTI 58 328 329 354 109 109 109 109TCRASQSVSSAVAWYQQKPGK APKLLIYSASSLYSGVPSRFSGS RSGTDFTLTISSLQPEDFATYYCQQSSQYDLITFGQGTKVEIKR G9.2-19 V_(H): EVQLVESGGGLVQPGGSLR 59 430 460 409160 213 266 319 LSCAASGFTVSSYSIHWVRQAP GKGLEWVASISSSSGSTSYADSVKGRFTISADTSKNTAYLQMNS LRAEDTAVYYCARNAWHYEPS YWYGNYATYGFDYWGQGTLV TVSSG9.2-20 V_(L): DIQMTQSPSSLSASVGDRVTI 54 328 329 352 108 108 108 108TCRASQSVSSAVAWYQQKPGK APKLLIYSASSLYSGVPSRFSGS RSGTDFTLTISSLQPEDFATYYCQQSSTDPITFGQGTKVEIKR G9.2-20 V_(H): EVQLVESGGGLVQPGGSLR 60 429 461 410161 214 267 320 LSCAASGFTVSYSSIHWVRQAP GKGLEWVASISSSSSSTYYADSVKGRFTISADTSKNTAYLQMNS LRAEDTAVYYCARGQQYYPD QYWGLDYWGQGTLVTVSS G9.2-21V_(L): DIQMTQSPSSLSASVGDRVTI 61 328 329 355 110 110 110 110TCRASQSVSSAVAWYQQKPGK APKLLIYSASSLYSGVPSRFSGS RSGTDFTLTISSLQPEDFATYYCQQSSSSSLFTFGQGTKVEIKR G9.2-21 V_(H): EVQLVESGGGLVQPGGSLR 62 429 462 411162 215 268 321 LSCAASGFTVSYSSIHWVRQAP GKGLEWVASIYSSSGYTYYADSVKGRFTISADTSKNTAYLQMN SLRAEDTAVYYCARTYYTYFD WWRTAVYYGFDYWGQGTLV TVSSG9.2-22 V_(L): DIQMTQSPSSLSASVGDRVTI 63 328 329 356 111 111 111 111TCRASQSVSSAVAWYQQKPGK APKLLIYSASSLYSGVPSRFSGS RSGTDFTLTISSLQPEDFATYYCQQRWYPGDLITFGQGTKVEIK R G9.2-22 V_(H): EVQLVESGGGLVQPGGSLR 64 428 463412 163 216 269 322 LSCAASGFTVYSSSIHWVRQAP GKGLEWVASISSSYGYTSYADSVKGRFTISADTSKNTAYLQMNS LRAEDTAVYYCARDYYNYMSS YWWYSALDYWGQGTLVTVSSG9.2-23 V_(L): DIQMTQSPSSLSASVGDRVTI 65 328 329 357 112 112 112 112TCRASQSVSSAVAWYQQKPGK APKLLIYSASSLYSGVPSRFSGS RSGTDFTLTISSLQPEDFATYYCQQSYFPSLVTFGQGTKVEIKR G9.2-23 V_(H): EVQLVESGGGLVQPGGSLR 66 428 464 413164 217 270 323 LSCAASGFTVYSSSIHWVRQAP GKGLEWVASIYPYYGYTSYADSVKGRFTISADTSKNTAYLQMN SLRAEDTAVYYCARKIFWPVS WMWQGYYPALDYWGQGTLV TVSSG9.2-24 V_(L): DIQMTQSPSSLSASVGDRVTI 67 328 329 358 113 113 113 113TCRASQSVSSAVAWYQQKPGK APKLLIYSASSLYSGVPSRFSGS RSGTDFTLTISSLQPEDFATYYCQQWSQSPVTFGQGTKVEIKR G9.2-24 V_(H): EVQLVESGGGLVQPGGSLR 68 428 465 414165 218 271 324 LSCAASGFTVYSSSIHWVRQAP GKGLEWVASIYSSYGYTSYADSVKGRFTISADTSKNTAYLQMN SLRAEDTAVYYCARSYSSETHY GWAMDYWGQGTLVTVSS G9.2-25V_(L): DIQMTQSPSSLSASVGDRVTI 69 328 329 359 114 114 114 114TCRASQSVSSAVAWYQQKPGK APKLLIYSASSLYSGVPSRFSGS RSGTDFTLTISSLQPEDFATYYCQQSYVYYPFTFGQGTKVEIKR G9.2-25 V_(H): EVQLVESGGGLVQPGGSLR 70 427 466 415166 219 272 325 LSCAASGFTLSSSSIHWVRQAP GKGLEWVASIYSSYGSTSYADSVKGRFTISADTSKNTAYLQMNS LRAEDTAVYYCARQYYTYFE WYMGWGYALDYWGQGTLVT VSSG9.2-26 V_(L): DIQMTQSPSSLSASVGDRVTI 71 328 329 360 115 115 115 115TCRASQSVSSAVAWYQQKPGK APKLLIYSASSLYSGVPSRFSGS RSGTDFTLTISSLQPEDFATYYCQQGGWYYGPITFGQGTKVEIK R G9.2-26 V_(H): EVQLVESGGGLVQPGGSLR 72 361 389416 167 220 273 326 LSCAASGFTVSSSSIHWVRQAP GKGLEWVAYISSYSGSTYYADSVKGRFTISADTSKNTAYLQMN SLRAEDTAVYYCARSSALYWM DFSYSALDYWGQGTLVTVSS G9.2-V_(L): DIQMTQSPSSLSASVGDRVTI 54 328 329 352 108 108 108 108 lowTCRASQSVSSAVAWYQQKPGK affinity APKLLIYSASSLYSGVPSRFSGS binderRSGTDFTLTISSLQPEDFATYYC QQSSTDPITFGQGTKVEIKR G9.2- V_(H):EVQLVESGGGLVQPGGSLR 73 361 388 417 168 221 274 327 lowLSCAASGFTVSSSSIHWVRQAP affinity GKGLEWVAYISSSSGYTYYAD binderSVKGRFTISADTSKNTAYLQMN SLRAEDTAVYYCARSSSSSSSSS SSSSDYWGQGTLVTVSS

TABLE 3 Selected Antibody CDR Sequences Clone Sequence SEQ ID NO: G9.1-8V_(L) CDR1 RASQSVSSAVA 328 V_(L) CDR2 SASSLYS 329 V_(L) CDR3 QQSYYDSNPIT337 V_(L) CDR1 FTVSSSSIH 361 V_(H) CDR2 YIYPYSGYTSYADSVKG 364 V_(H) CDR3YSTYSWGGIGKWVWGMDY 374 G9.1-8m1 V_(L) CDR1 RASQSVSSAVA 328 V_(L) CDR2SASSLYS 329 V_(L) CDR3 QQSYYDSNPIT 337 V_(L) CDR1 FTVSSSSIH 361 V_(H)CDR2 SSSSSSGYTSYADSVKG 365 V_(H) CDR3 YSTYSWGGIGKWVWGMDY 374 G9.1-8m2V_(L) CDR1 RASQSVSSAVA 328 V_(L) CDR2 SASSLYS 329 V_(L) CDR3 QQSYYDSNPIT337 V_(L) CDR1 FTVSSSSIH 361 V_(H) CDR2 YIYPYSSSSSYADSVKG 366 V_(H) CDR3YSTYSWGGIGKWVWGMDY 374 G9.1-8m3 V_(L) CDR1 RASQSVSSAVA 328 V_(L) CDR2SASSLYS 329 V_(L) CDR3 QQSYYDSNPIT 337 V_(L) CDR1 FTVSSSSIH 361 V_(H)CDR2 YIYPYSGYTSYADSVKG 364 V_(H) CDR3 SSSSSWGGIGKWVWGMDY 375 G9.1-8m4V_(L) CDR1 RASQSVSSAVA 328 V_(L) CDR2 SASSLYS 329 V_(L) CDR3 QQSYYDSNPIT337 V_(L) CDR1 FTVSSSSIH 361 V_(H) CDR2 YIYPYSGYTSYADSVKG 364 V_(H) CDR3YSTYSSSSSSKWVWGMDY 376 G9.1-8m5 V_(L) CDR1 RASQSVSSAVA 328 V_(L) CDR2SASSLYS 329 V_(L) CDR3 QQSYYDSNPIT 337 V_(H) CDR1 FTVSSSSIH 361 V_(H)CDR2 YIYPYSGYTSYADSVKG 364 V_(H) CDR3 YSTYSWGGIGSSSSSMDY 377 G9.1-8m6V_(L) CDR1 RASQSVSSAVA 328 V_(L) CDR2 SASSLYS 329 V_(L) CDR3 QQSYYDSNPIT337 V_(H) CDR1 FTVSSSSIH 361 V_(H) CDR2 YIYPYSGYTSYADSVKG 364 V_(H) CDR3YSTYSSSSSKWVWGMDY 378 G9.1-8m7 V_(L) CDR1 RASQSVSSAVA 328 V_(L) CDR2SASSLYS 329 V_(L) CDR3 QQSYYDSNPIT 337 V_(H) CDR1 FTVSSSSIH 361 V_(H)CDR2 YIYPYSGYTSYADSVKG 364 V_(H) CDR3 YSTYSSSSKWVWGMDY 379 G9.1-8m8V_(L) CDR1 RASQSVSSAVA 328 V_(L) CDR2 SASSLYS 329 V_(L) CDR3 QQSYYDSNPIT337 V_(H) CDR1 FTVSSSSIH 361 V_(H) CDR2 YIYPYSGYTSYADSVKG 364 V_(H) CDR3YSTYSSSKWVWGMDY 380 G9.1-8m9 V_(L) CDR1 RASQSVSSAVA 328 V_(L) CDR2SASSLYS 329 V_(L) CDR3 QQSYYDSNPIT 337 V_(H) CDR1 FTVSSSSIH 361 V_(H)CDR2 YIYPYSGYTSYADSVKG 364 V_(H) CDR3 YSTYSSKWVWGMDY 383 G9.1-8m10 V_(L)CDR1 RASQSVSSAVA 328 V_(L) CDR2 SASSLYS 329 V_(L) CDR3 QQSYYDSNPIT 337V_(H) CDR1 FTVSSSSIH 361 V_(H) CDR2 YIYPYSGYTSYADSVKG 364 V_(H) CDR3YSTYSKWVWGMDY 381 G9.1-8m11 V_(L) CDR1 RASQSVSSAVA 328 V_(L) CDR2SASSLYS 329 V_(L) CDR3 QQSYYDSNPIT 337 V_(H) CDR1 FTVSSSSIH 361 V_(H)CDR2 YIYPYSGYTSYADSVKG 364 V_(H) CDR3 YSTYKWVWGMDY 382 G9.1-8m12 V_(L)CDR1 RASQSVSSAVA 328 V_(L) CDR2 SASSLYS 329 V_(L) CDR3 QQSYYDSNPIT 337V_(H) CDR1 FTVSSSSIH 361 V_(H) CDR2 YIYPYSSSSSYADSVKG 366 V_(H) CDR3YSTYSSSKWVWGMDY 380 G9.1-8m13 V_(L) CDR1 RASQSVSSAVA 328 V_(L) CDR2SASSLYS 329 V_(L) CDR3 QQSYYDSNPIT 337 V_(H) CDR1 FTVSSSSIH 361 V_(H)CDR2 YIYPYSSSSSYADSVKG 366 V_(H) CDR3 YSTYSSKWVWGMDY 383 G9.1-8m14 V_(L)CDR1 RASQSVSSAVA 328 V_(L) CDR2 SASSLYS 329 V_(L) CDR3 QQSYYDSNPIT 337V_(H) CDR1 FTVSSSSIH 361 V_(H) CDR2 YIYPYSSSSSYADSVKG 366 V_(H) CDR3YSTYKWVWGMDY 382 G9.2-17 V_(L) CDR1 RASQSVSSAVA 328 V_(L) CDR2 SASSLYS329 V_(L) CDR3 QQSSTDPIT 352 V_(H) CDR1 FTVSSSSIH 361 V_(H) CDR2YISSSSGYTYYADSVKG 388 V_(H) CDR3 YWSYPSWWPYRGMDY 406 G9.2-17m6 V_(L)CDR1 RASQSVSSAVA 328 V_(L) CDR2 SASSLYS 329 V_(L) CDR3 QQSSTDPIT 352V_(H) CDR1 FTVSSSSIH 361 V_(H) CDR2 YISSSSGYTYYADSVKG 388 V_(H) CDR3YWSYPSWSPYRGMDY 407

Such CRD1 and CRD2 binding anti-Galectin-9 antibodies are isolated andstructurally characterized as described herein. The disclosure alsocontemplates antibodies having at least 80% identity (e.g., at least85%, at least 90%, at least 95%, or at least 99% identity) to theirvariable region or CDR sequences. The VL amino acid sequences of G9.2-1,G9.2-2, G9.2-3, G9.2-4, G9.2-5, G9.2-6, G9.2-7, G9.2-8, G9.2-9, G9.2-10,G9.2-11, G9.2-12, G9.2-13, G9.2-14, G9.2-15, G9.2-16, G9.2-17,G9.2-17mut6, G9.2-18, G9.2-19, G9.2-20, G9.2-21, G9.2-22, G9.2-23,G9.2-24, G9.2-25, G9.2-26, G9.2-low affinity binder are set forth in SEQID NO: 29, 13, 34, 36, 38, 40, 42, 44, 46, 48, 29, 34, 54, 58, 61, 63,65, 73, 67, 69, and 71. The VH amino acid sequences of G9.2-1, G9.2-2,G9.2-3, G9.2-4, G9.2-5, G9.2-6, G9.2-7, G9.2-8, G9.2-9, G9.2-10,G9.2-11, G9.2-12, G9.2-13, G9.2-14, G9.2-15, G9.2-16, G9.2-17,G9.2-17mut6, G9.2-18, G9.2-19, G9.2-20, G9.2-21, G9.2-22, G9.2-23,G9.2-24, G9.2-25, G9.2-26, G9.2-low affinity binder are set forth in SEQID NO: 30, 31, 32, 33, 35, 37, 39, 41, 43, 45, 47, 49, 50, 51, 52, 53,55, 56, 57, 59, 60, 62, 64, 66, 68, 70, 72 and 73. Accordingly, providedherein are isolated anti-Galectin-9 antibodies, or antigen bindingportion thereof, comprising heavy and light chain variable regions,wherein the light chain variable region comprises an amino acid sequenceselected from SEQ ID NO: 29, 13, 34, 36, 38, 40, 42, 44, 46, 48, 29, 34,54, 58, 61, 63, 65, 73, 67, 69, and 71. In some embodiments, the lightchain variable regions consists of an amino acid sequence selected fromSEQ ID NO: 29, 13, 34, 36, 38, 40, 42, 44, 46, 48, 29, 34, 54, 58, 61,63, 65, 73, 67, 69, and 71. Also provided are isolated anti-Galectin-9antibodies, or antigen binding portions thereof, comprising heavy andlight chain variable regions, wherein the heavy chain variable regioncomprises an amino acid sequence selected from SEQ ID NO: 30, 31, 32,33, 35, 37, 39, 41, 43, 45, 47, 49, 50, 51, 52, 53, 55, 56, 57, 59, 60,62, 64, 66, 68, 70, 72 and 73. In some embodiments, the heavy chainvariable regions consists of an amino acid sequence selected from SEQ IDNO: 30, 31, 32, 33, 35, 37, 39, 41, 43, 45, 47, 49, 50, 51, 52, 53, 55,56, 57, 59, 60, 62, 64, 66, 68, 70, 72 and 73. Accordingly, providedherein are isolated anti-Galectin-9 antibodies, or antigen bindingportion thereof, comprising heavy and light chain variable regions,wherein the light chain variable region comprises an amino acid sequenceselected from SEQ ID NO: 29, 13, 34, 36, 38, 40, 42, 44, 46, 48, 29, 34,54, 58, 61, 63, 65, 73, 67, 69, and 71, and the heavy chain variableregion comprises an amino acid sequence selected from SEQ ID NO: 30, 31,32, 33, 35, 37, 39, 41, 43, 45, 47, 49, 50, 51, 52, 53, 55, 56, 57, 59,60, 62, 64, 66, 68, 70, 72 and 73. Accordingly, provided herein areisolated anti-Galectin-9 antibodies, or antigen binding portion thereof,comprising heavy and light chain variable regions, wherein the lightchain variable region consists of an amino acid sequence selected fromSEQ ID NO: 29, 13, 34, 36, 38, 40, 42, 44, 46, 48, 29, 34, 54, 58, 61,63, 65, 73, 67, 69, and 71, and the heavy chain variable region consistsof an amino acid sequence selected from SEQ ID NO: 30, 31, 32, 33, 35,37, 39, 41, 43, 45, 47, 49, 50, 51, 52, 53, 55, 56, 57, 59, 60, 62, 64,66, 68, 70, 72 and 73.

In some embodiments, the anti-Galectin-9 antibody comprises a VL regionhaving the sequence of SEQ ID NO: 54. In some embodiments, theanti-Galectin-9 antibody comprises a VH region having the sequence ofSEQ ID NO: 55. In some embodiments, the anti-Galectin-9 antibodycomprises a VH region having the sequence of SEQ ID NO: 56. In someembodiments, the anti-Galectin-9 antibody comprises a VL region havingthe sequence of SEQ ID NO: 54 and a VH region having the sequence of SEQID NO: 55. In some embodiments, the anti-Galectin-9 antibody comprises aVL region having the sequence of SEQ ID NO: 54 and a VH region havingthe sequence of SEQ ID NO: 56.

The VL amino acid sequences of G9.1-1, G9.1-2, G9.1-3, G9.1-4, G9.1-5,G9.1-6, G9.1-7, G9.1-8, G9.1-9, G9.1-10, G9.1-11, G9.1-8m1, G9.1-8m2,G9.1-8m3, G9.1-8m4, G9.1-8m5, G9.1-8m6, G9.1-8m7, G9.1-8m8, G9.1-8m9,G9.1-8m10, G9.1-8m11, G9.1-8m12, G9.1-8m13, G9.1-8m14 are set forth inSEQ ID NO: 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, and 27, respectively.The VH amino acid sequences of G9.1-1, G9.1-2, G9.1-3, G9.1-4, G9.1-5,G9.1-6, G9.1-7, G9.1-8, G9.1-9, G9.1-10, G9.1-11, G9.1-8m1, G9.1-8m2,G9.1-8m3, G9.1-8m4, G9.1-8m5, G9.1-8m6, G9.1-8m7, G9.1-8m8, G9.1-8m9,G9.1-8m10, G9.1-8m11, G9.1-8m12, G9.1-8m13, G9.1-8m14 are set forth inSEQ ID NO: 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 74, 75, 76, 77,78, 79, 80, 81, 82, 83, 84, 85, 86, and 87. Accordingly, provided hereinare isolated anti-Galectin-9 antibodies, or antigen binding portionsthereof, comprising heavy and light chain variable regions, wherein thelight chain variable region comprises an amino acid sequence selectedfrom SEQ ID NO: 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, and 27. In someembodiments, the light chain variable region consists of an amino acidsequence selected from SEQ ID NO: 7, 9, 11, 13, 15, 17, 19, 21, 23, 25,and 27. Also provided are isolated anti-Galectin-9 antibodies, orantigen binding portions thereof, comprising heavy and light chainvariable regions, wherein the heavy chain variable region comprises anamino acid sequence selected from SEQ ID NO: 8, 10, 12, 14, 16, 18, 20,22, 24, 26, 28, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, and87. In some embodiments, the heavy chain variable regions consists of anamino acid sequence selected from SEQ ID NO: 8, 10, 12, 14, 16, 18, 20,22, 24, 26, 28, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, and87. Accordingly, provided herein are isolated anti-Galectin-9antibodies, or antigen binding portions thereof, comprising heavy andlight chain variable regions, wherein the light chain variable regioncomprises an amino acid sequence selected from SEQ ID NO: 7, 9, 11, 13,15, 17, 19, 21, 23, 25, and 27 and the heavy chain variable regioncomprises an amino acid sequence selected from SEQ ID NO: 8, 10, 12, 14,16, 18, 20, 22, 24, 26, 28, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84,85, 86, and 87. In some embodiments, the light chain variable regionsconsists of an amino acid sequence selected from SEQ ID NO: 7, 9, 11,13, 15, 17, 19, 21, 23, 25, and 27, and the heavy chain variable regionsconsists of an amino acid sequence selected from SEQ ID NO: 8, 10, 12,14, 16, 18, 20, 22, 24, 26, 28, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83,84, 85, 86, and 87.

In some embodiments, the anti-Galectin-9 antibody comprises a VL regionhaving the sequence of SEQ ID NO: 21. In some embodiments, theanti-Galectin-9 antibody comprises a VH region having the sequence ofSEQ ID NO: 22. In some embodiments, the anti-Galectin-9 antibodycomprises a VH region having the sequence of SEQ ID NO: 86. In someembodiments, the anti-Galectin-9 antibody comprises a VL region havingthe sequence of SEQ ID NO: 21 and a VH region having the sequence of SEQID NO: 22. In some embodiments, the anti-Galectin-9 antibody comprises aVL region having the sequence of SEQ ID NO: 21 and a VH region havingthe sequence of SEQ ID NO: 86.

In some specific embodiments, the anti-Galectin-9 antibody or antigenbinding portion thereof comprises any of SEQ ID NO: 7-87. In somespecific embodiments, the anti-Galectin-9 antibody comprises one or moresequences of any sequence(s) selected from SEQ ID NO: 7-87 and anycombination(s) thereof.

In some specific embodiments, the anti-Galectin-9 antibody or antigenbinding portion thereof comprises any of SEQ ID NOs: 7-28 and 74-87. Insome specific embodiments, the anti-Galectin-9 antibody or antigenbinding portion thereof comprises one or more sequences of anysequence(s) selected from SEQ ID NO: 7-28 and 74-87 and anycombination(s) thereof.

In some specific embodiments, the anti-Galectin-9 antibody or antigenbinding portion thereof comprises any of SEQ ID NOs: 13, 29-73. In somespecific embodiments, the anti-Galectin-9 antibody or antigen bindingportion thereof comprises one or more sequences of any sequence(s)selected from SEQ ID NO: 13, 29-73 and any combination(s) thereof. Insome specific embodiments, the anti-Galectin-9 antibody or antigenbinding portion thereof comprises any of SEQ ID NOs: 54, 55, or 54 and56. In some specific embodiments, the anti-Galectin-9 antibody orantigen binding portion thereof comprises one or more sequences of anysequence(s) selected from SEQ ID NO: 54, 55, or 54 and 56 and anycombination(s) thereof.

In some embodiments, the anti-Galectin-9 antibody or antigen bindingportion thereof comprises a VL region comprising SEQ ID NO: 54. In someembodiments, the anti-Galectin-9 antibody or antigen binding portionthereof comprises a VH region comprising SEQ ID NO: 55. In someembodiments, the anti-Galectin-9 antibody comprises a VL regionconsisting of SEQ ID NO: 54. In some embodiments, the anti-Galectin-9antibody or antigen binding portion thereof comprises a VH regionconsisting of SEQ ID NO: 55. In some embodiments, the anti-Galectin-9antibody comprises a VL and VH region comprising SEQ ID NO: 54 and 55.In some specific embodiments, the anti-Galectin-9 antibody or antigenbinding portion thereof comprises a VL and VH region consisting of SEQID NO: 54 and 55.

In some embodiments, the anti-Galectin-9 antibody or antigen bindingportion thereof comprises a VL region comprising SEQ ID NO: 21. In someembodiments, the anti-Galectin-9 antibody or antigen binding portionthereof comprises a VL region consisting of SEQ ID NO: 21. In someembodiments, the anti-Galectin-9 antibody or antigen binding portionthereof comprises a VH region comprising SEQ ID NO: 86. In someembodiments, the anti-Galectin-9 antibody or antigen binding portionthereof comprises a VH region consisting of SEQ ID NO: 86. In someembodiments, the anti-Galectin-9 antibody or antigen binding portionthereof comprises a VL and VH region comprising SEQ ID NO: 21 and 86. Insome embodiments, the anti-Galectin-9 antibody or antigen bindingportion thereof comprises a VL and VH region consisting of SEQ ID NO: 21and 86.

In some specific embodiments, the anti-Galectin-9 antibody or antigenbinding portion thereof comprises a VL or VH region comprising any of ofSEQ ID NOs: 21, 22 and 74-87. In some specific embodiments, theanti-Galectin-9 antibody or antigen binding portion thereof comprises aVL or VH region consisting of SEQ ID NOs: 21, 22 and 74-87. In somespecific embodiments, the anti-Galectin-9 antibody or antigen bindingportion thereof comprises a VL and/or VH region comprising sequence(s)selected from SEQ ID NO: 21, 22 and 74-87 and any combination(s)thereof. In some specific embodiments, the anti-Galectin-9 antibody orantigen binding portion thereof comprises a VL and/or VH regionconsisting of sequence(s) selected from SEQ ID NO: 21, 22 and 74-87 andany combination(s) thereof.

In some embodiments, the anti-Galectin-9 antibody or antigen bindingportion thereof comprises a VL region and a VH region comprising SEQ IDNO: 7 and SEQ ID NO: 8. In some embodiments, the anti-Galectin-9antibody or antigen binding portion thereof comprises a VL region and aVH region comprising SEQ ID NO: 9 and SEQ ID NO: 10. In someembodiments, the anti-Galectin-9 antibody or antigen binding portionthereof comprises a VL region and a VH region comprising SEQ ID NO: 11and SEQ ID NO: 12. In some embodiments, the anti-Galectin-9 antibody orantigen binding portion thereof comprises a VL region and a VH regioncomprising SEQ ID NO: 13 and SEQ ID NO: 14. In some embodiments, theanti-Galectin-9 antibody or antigen binding portion thereof comprises aVL region and a VH region comprising SEQ ID NO: 15 and SEQ ID NO: 16. Insome embodiments, the anti-Galectin-9 antibody or antigen bindingportion thereof comprises a VL region and a VH region comprising SEQ IDNO: 17 and SEQ ID NO: 18. In some embodiments, the anti-Galectin-9antibody or antigen binding portion thereof comprises a VL region and aVH region comprising SEQ ID NO: 19 and SEQ ID NO: 20. In someembodiments, the anti-Galectin-9 antibody or antigen binding portionthereof comprises a VL region and a VH region comprising SEQ ID NO: 21and SEQ ID NO: 22. In some embodiments, the anti-Galectin-9 antibody orantigen binding portion thereof comprises a VL region and a VH regioncomprising SEQ ID NO: 23 and SEQ ID NO: 24. In some embodiments, theanti-Galectin-9 antibody or antigen binding portion thereof comprises aVL region and a VH region comprising SEQ ID NO: 25 and SEQ ID NO: 26. Insome embodiments, the anti-Galectin-9 antibody or antigen bindingportion thereof comprises a VL region and a VH region comprising SEQ IDNO: 27 and SEQ ID NO: 28. In some embodiments, the anti-Galectin-9antibody or antigen binding portion thereof comprises a VL region and aVH region comprising SEQ ID NO: 21 and SEQ ID NO: 74. In someembodiments, the anti-Galectin-9 antibody or antigen binding portionthereof comprises a VL region and a VH region comprising SEQ ID NO: 21and SEQ ID NO: 75. In some embodiments, the anti-Galectin-9 antibody orantigen binding portion thereof comprises a VL region and a VH regioncomprising SEQ ID NO: 21 and SEQ ID NO: 76. In some embodiments, theanti-Galectin-9 antibody or antigen binding portion thereof comprises aVL region and a VH region comprising SEQ ID NO: 21 and SEQ ID NO: 77. Insome embodiments, the anti-Galectin-9 antibody or antigen bindingportion thereof comprises a VL region and a VH region comprising SEQ IDNO: 21 and SEQ ID NO: 78. In some embodiments, the anti-Galectin-9antibody or antigen binding portion thereof comprises a VL region and aVH region comprising SEQ ID NO: 21 and SEQ ID NO: 79. In someembodiments, the anti-Galectin-9 antibody or antigen binding portionthereof comprises a VL region and a VH region comprising SEQ ID NO: 21and SEQ ID NO: 80. In some embodiments, the anti-Galectin-9 antibody orantigen binding portion thereof comprises a VL region and a VH regioncomprising SEQ ID NO: 21 and SEQ ID NO: 81. In some embodiments, theanti-Galectin-9 antibody or antigen binding portion thereof comprises aVL region and a VH region comprising SEQ ID NO: 21 and SEQ ID NO: 82. Insome embodiments, the anti-Galectin-9 antibody or antigen bindingportion thereof comprises a VL region and a VH region comprising SEQ IDNO: 21 and SEQ ID NO: 83. In some embodiments, the anti-Galectin-9antibody or antigen binding portion thereof comprises a VL region and aVH region comprising SEQ ID NO: 21 and SEQ ID NO: 84. In someembodiments, the anti-Galectin-9 antibody or antigen binding portionthereof comprises a VL region and a VH region comprising SEQ ID NO: 108and SEQ ID NO: 85. In some embodiments, the anti-Galectin-9 antibody orantigen binding portion thereof comprises a VL region and a VH regioncomprising SEQ ID NO: 21 and SEQ ID NO: 86. In some embodiments, theanti-Galectin-9 antibody or antigen binding portion thereof comprises aVL region and a VH region comprising SEQ ID NO: 29 and SEQ ID NO: 30. Insome embodiments, the anti-Galectin-9 antibody or antigen bindingportion thereof comprises a VL region and a VH region comprising SEQ IDNO: 13 and SEQ ID NO: 31. In some embodiments, the anti-Galectin-9antibody or antigen binding portion thereof comprises a VL region and aVH region comprising SEQ ID NO: 13 and SEQ ID NO: 32. In someembodiments, the anti-Galectin-9 antibody or antigen binding portionthereof comprises a VL region and a VH region comprising SEQ ID NO: 13and SEQ ID NO: 33. In some embodiments, the anti-Galectin-9 antibody orantigen binding portion thereof comprises a VL region and a VH regioncomprising SEQ ID NO: 34 and SEQ ID NO: 35. In some embodiments, theanti-Galectin-9 antibody or antigen binding portion thereof comprises aVL region and a VH region comprising SEQ ID NO: 36 and SEQ ID NO: 37. Insome embodiments, the anti-Galectin-9 antibody or antigen bindingportion thereof comprises a VL region and a VH region comprising SEQ IDNO: 38 and SEQ ID NO: 39. In some embodiments, the anti-Galectin-9antibody or antigen binding portion thereof comprises a VL region and aVH region comprising SEQ ID NO: 40 and SEQ ID NO: 41. In someembodiments, the anti-Galectin-9 antibody or antigen binding portionthereof comprises a VL region and a VH region comprising SEQ ID NO: 42and SEQ ID NO: 43. In some embodiments, the anti-Galectin-9 antibody orantigen binding portion thereof comprises a VL region and a VH regioncomprising SEQ ID NO: 44 and SEQ ID NO: 45. In some embodiments, theanti-Galectin-9 antibody or antigen binding portion thereof comprises aVL region and a VH region comprising SEQ ID NO: 46 and SEQ ID NO: 47. Insome embodiments, the anti-Galectin-9 antibody or antigen bindingportion thereof comprises a VL region and a VH region comprising SEQ IDNO: 48 and SEQ ID NO: 49. In some embodiments, the anti-Galectin-9antibody or antigen binding portion thereof comprises a VL region and aVH region comprising SEQ ID NO: 29 and SEQ ID NO: 50. In someembodiments, the anti-Galectin-9 antibody or antigen binding portionthereof comprises a VL region and a VH region comprising SEQ ID NO: 13and SEQ ID NO: 51. In some embodiments, the anti-Galectin-9 antibody orantigen binding portion thereof comprises a VL region and a VH regioncomprising SEQ ID NO: 34 and SEQ ID NO: 52. In some embodiments, theanti-Galectin-9 antibody or antigen binding portion thereof comprises aVL region and a VH region comprising SEQ ID NO: 13 and SEQ ID NO: 53. Insome embodiments, the anti-Galectin-9 antibody or antigen bindingportion thereof comprises a VL region and a VH region comprising SEQ IDNO: 54 and SEQ ID NO: 55. In some embodiments, the anti-Galectin-9antibody or antigen binding portion thereof comprises a VL region and aVH region comprising SEQ ID NO: 13 and SEQ ID NO: 57. In someembodiments, the anti-Galectin-9 antibody or antigen binding portionthereof comprises a VL region and a VH region comprising SEQ ID NO: 58and SEQ ID NO: 59. In some embodiments, the anti-Galectin-9 antibody orantigen binding portion thereof comprises a VL region and a VH regioncomprising SEQ ID NO: 54 and SEQ ID NO: 60. In some embodiments, theanti-Galectin-9 antibody or antigen binding portion thereof comprises aVL region and a VH region comprising SEQ ID NO: 61 and SEQ ID NO: 62. Insome embodiments, the anti-Galectin-9 antibody or antigen bindingportion thereof comprises a VL region and a VH region comprising SEQ IDNO: 63 and SEQ ID NO: 64. In some embodiments, the anti-Galectin-9antibody or antigen binding portion thereof comprises a VL region and aVH region comprising SEQ ID NO: 65 and SEQ ID NO: 66. In someembodiments, the anti-Galectin-9 antibody or antigen binding portionthereof comprises a VL region and a VH region comprising SEQ ID NO: 54and SEQ ID NO: 56. In some embodiments, the anti-Galectin-9 antibody orantigen binding portion thereof comprises a VL region and a VH regioncomprising SEQ ID NO: 67 and SEQ ID NO: 68. In some embodiments, theanti-Galectin-9 antibody or antigen binding portion thereof comprises aVL region and a VH region comprising SEQ ID NO: 69 and SEQ ID NO: 70. Insome embodiments, the anti-Galectin-9 antibody or antigen bindingportion thereof comprises a VL region and a VH region comprising SEQ IDNO: 71 and SEQ ID NO: 72. In some embodiments, the anti-Galectin-9antibody or antigen binding portion thereof comprises a VL region and aVH region comprising SEQ ID NO: 54 and SEQ ID NO: 73. In some specificembodiments, the anti-Galectin-9 antibody or antigen binding portionthereof comprises a VL region and a VH region consisting of SEQ ID NO: 7and SEQ ID NO: 8. In some specific embodiments, the anti-Galectin-9antibody or antigen binding portion thereof comprises a VL region and aVH region consisting of SEQ ID NO: 9 and SEQ ID NO: 10. In some specificembodiments, the anti-Galectin-9 antibody or antigen binding portionthereof comprises a VL region and a VH region consisting of SEQ ID NO:11 and SEQ ID NO: 12. In some specific embodiments, the anti-Galectin-9antibody or antigen binding portion thereof comprises a VL region and aVH region consisting of SEQ ID NO: 13 and SEQ ID NO: 14. In somespecific embodiments, the anti-Galectin-9 antibody or antigen bindingportion thereof comprises a VL region and a VH region consisting of SEQID NO: 15 and SEQ ID NO: 16. In some specific embodiments, theanti-Galectin-9 antibody or antigen binding portion thereof comprises aVL region and a VH region consisting of SEQ ID NO: 17 and SEQ ID NO: 18.In some specific embodiments, the anti-Galectin-9 antibody or antigenbinding portion thereof comprises a VL region and a VH region consistingof SEQ ID NO: 19 and SEQ ID NO: 20. In some specific embodiments, theanti-Galectin-9 antibody or antigen binding portion thereof comprises aVL region and a VH region consisting of SEQ ID NO: 21 and SEQ ID NO: 22.In some specific embodiments, the anti-Galectin-9 antibody or antigenbinding portion thereof comprises a VL region and a VH region consistingof SEQ ID NO: 23 and SEQ ID NO: 24. In some specific embodiments, theanti-Galectin-9 antibody or antigen binding portion thereof comprises aVL region and a VH region consisting of SEQ ID NO: 25 and SEQ ID NO: 26.In some specific embodiments, the anti-Galectin-9 antibody or antigenbinding portion thereof comprises a VL region and a VH region consistingof SEQ ID NO: 27 and SEQ ID NO: 28. In some specific embodiments, theanti-Galectin-9 antibody or antigen binding portion thereof comprises aVL region and a VH region consisting of SEQ ID NO: 21 and SEQ ID NO: 74.In some specific embodiments, the anti-Galectin-9 antibody or antigenbinding portion thereof comprises a VL region and a VH region consistingof SEQ ID NO: 21 and SEQ ID NO: 75. In some specific embodiments, theanti-Galectin-9 antibody or antigen binding portion thereof comprises aVL region and a VH region consisting of SEQ ID NO: 21 and SEQ ID NO: 76.In some specific embodiments, the anti-Galectin-9 antibody or antigenbinding portion thereof comprises a VL region and a VH region consistingof SEQ ID NO: 21 and SEQ ID NO: 77. In some specific embodiments, theanti-Galectin-9 antibody or antigen binding portion thereof comprises aVL region and a VH region consisting of SEQ ID NO: 21 and SEQ ID NO: 78.In some specific embodiments, the anti-Galectin-9 antibody or antigenbinding portion thereof comprises a VL region and a VH region consistingof SEQ ID NO: 21 and SEQ ID NO: 79. In some specific embodiments, theanti-Galectin-9 antibody or antigen binding portion thereof comprises aVL region and a VH region consisting of SEQ ID NO: 21 and SEQ ID NO: 80.In some specific embodiments, the anti-Galectin-9 antibody or antigenbinding portion thereof comprises a VL region and a VH region consistingof SEQ ID NO: 21 and SEQ ID NO: 81. In some specific embodiments, theanti-Galectin-9 antibody or antigen binding portion thereof comprises aVL region and a VH region consisting of SEQ ID NO: 21 and SEQ ID NO: 82.In some specific embodiments, the anti-Galectin-9 antibody or antigenbinding portion thereof comprises a VL region and a VH region consistingof SEQ ID NO: 21 and SEQ ID NO: 83. In some specific embodiments, theanti-Galectin-9 antibody or antigen binding portion thereof comprises aVL region and a VH region consisting of SEQ ID NO: 21 and SEQ ID NO: 84.In some specific embodiments, the anti-Galectin-9 antibody or antigenbinding portion thereof comprises a VL region and a VH region consistingof SEQ ID NO: 108 and SEQ ID NO: 85. In some specific embodiments, theanti-Galectin-9 antibody or antigen binding portion thereof comprises aVL region and a VH region consisting of SEQ ID NO: 21 and SEQ ID NO: 86.In some specific embodiments, the anti-Galectin-9 antibody or antigenbinding portion thereof comprises a VL region and a VH region consistingof SEQ ID NO: 29 and SEQ ID NO: 30. In some specific embodiments, theanti-Galectin-9 antibody or antigen binding portion thereof comprises aVL region and a VH region consisting of SEQ ID NO: 13 and SEQ ID NO: 31.In some specific embodiments, the anti-Galectin-9 antibody or antigenbinding portion thereof comprises a VL region and a VH region consistingof SEQ ID NO: 13 and SEQ ID NO: 32. In some specific embodiments, theanti-Galectin-9 antibody or antigen binding portion thereof comprises aVL region and a VH region consisting of SEQ ID NO: 13 and SEQ ID NO: 33.In some specific embodiments, the anti-Galectin-9 antibody or antigenbinding portion thereof comprises a VL region and a VH region consistingof SEQ ID NO: 34 and SEQ ID NO: 35. In some specific embodiments, theanti-Galectin-9 antibody or antigen binding portion thereof comprises aVL region and a VH region consisting of SEQ ID NO: 36 and SEQ ID NO: 37.In some specific embodiments, the anti-Galectin-9 antibody or antigenbinding portion thereof comprises a VL region and a VH region consistingof SEQ ID NO: 38 and SEQ ID NO: 39. In some specific embodiments, theanti-Galectin-9 antibody or antigen binding portion thereof comprises aVL region and a VH region consisting of SEQ ID NO: 40 and SEQ ID NO: 41.In some specific embodiments, the anti-Galectin-9 antibody or antigenbinding portion thereof comprises a VL region and a VH region consistingof SEQ ID NO: 42 and SEQ ID NO: 43. In some specific embodiments, theanti-Galectin-9 antibody or antigen binding portion thereof comprises aVL region and a VH region consisting of SEQ ID NO: 44 and SEQ ID NO: 45.In some specific embodiments, the anti-Galectin-9 antibody or antigenbinding portion thereof comprises a VL region and a VH region consistingof SEQ ID NO: 46 and SEQ ID NO: 47. In some specific embodiments, theanti-Galectin-9 antibody or antigen binding portion thereof comprises aVL region and a VH region consisting of SEQ ID NO: 48 and SEQ ID NO: 49.In some specific embodiments, the anti-Galectin-9 antibody or antigenbinding portion thereof comprises a VL region and a VH region consistingof SEQ ID NO: 29 and SEQ ID NO: 50. In some specific embodiments, theanti-Galectin-9 antibody or antigen binding portion thereof comprises aVL region and a VH region consisting of SEQ ID NO: 13 and SEQ ID NO: 51.In some specific embodiments, the anti-Galectin-9 antibody or antigenbinding portion thereof comprises a VL region and a VH region consistingof SEQ ID NO: 34 and SEQ ID NO: 52. In some specific embodiments, theanti-Galectin-9 antibody or antigen binding portion thereof comprises aVL region and a VH region consisting of SEQ ID NO: 13 and SEQ ID NO: 53.In some specific embodiments, the anti-Galectin-9 antibody or antigenbinding portion thereof comprises a VL region and a VH region consistingof SEQ ID NO: 54 and SEQ ID NO: 55. In some specific embodiments, theanti-Galectin-9 antibody or antigen binding portion thereof comprises aVL region and a VH region consisting of SEQ ID NO: 13 and SEQ ID NO: 57.In some specific embodiments, the anti-Galectin-9 antibody or antigenbinding portion thereof comprises a VL region and a VH region consistingof SEQ ID NO: 58 and SEQ ID NO: 59. In some specific embodiments, theanti-Galectin-9 antibody or antigen binding portion thereof comprises aVL region and a VH region consisting of SEQ ID NO: 54 and SEQ ID NO: 60.In some specific embodiments, the anti-Galectin-9 antibody or antigenbinding portion thereof comprises a VL region and a VH region consistingof SEQ ID NO: 61 and SEQ ID NO: 62. In some specific embodiments, theanti-Galectin-9 antibody or antigen binding portion thereof comprises aVL region and a VH region consisting of SEQ ID NO: 63 and SEQ ID NO: 64.In some specific embodiments, the anti-Galectin-9 antibody or antigenbinding portion thereof comprises a VL region and a VH region consistingof SEQ ID NO: 65 and SEQ ID NO: 66. In some specific embodiments, theanti-Galectin-9 antibody or antigen binding portion thereof comprises aVL region and a VH region consisting of SEQ ID NO: 54 and SEQ ID NO: 56.In some specific embodiments, the anti-Galectin-9 antibody or antigenbinding portion thereof comprises a VL region and a VH region consistingof SEQ ID NO: 67 and SEQ ID NO: 68. In some specific embodiments, theanti-Galectin-9 antibody or antigen binding portion thereof comprises aVL region and a VH region consisting of SEQ ID NO: 69 and SEQ ID NO: 70.In some specific embodiments, the anti-Galectin-9 antibody or antigenbinding portion thereof comprises a VL region and a VH region consistingof SEQ ID NO: 71 and SEQ ID NO: 72. In some specific embodiments, theanti-Galectin-9 antibody or antigen binding portion thereof comprises aVL region and a VH region consisting of SEQ ID NO: 54 and SEQ ID NO: 73.

In some embodiments, the anti-Galectin-9 antibody comprises sequencehaving at least 80% (e.g., 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% andany incremental percent therein) sequence identity with any of theanti-Galectin-9 antibodies described in the previous paragraphs. In someembodiments, the anti-Galectin-9 antibody comprises a VL region that hasat least 80% (e.g., 85%, 90%, 95%, 96%, 97%, 98%, 99% and any incrementtherein) sequence identity to a VL region set forth in SEQ ID NOs: 7, 9,11, 13, 15, 17, 19, 21, 23, 25, and 27. In some embodiments, theanti-Galectin-9 antibody comprises a VH region that has at least 80%(e.g., 85%, 90%, 95%, 96%, 97%, 98%, 99% and any increment therein)sequence identity to a VH region set forth in SEQ ID NOs: 8, 10, 12, 14,16, 18, 20, 22, 24, 26, 28, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84,85, 86, and 87. In some embodiments, the anti-Galectin-9 antibodycomprises a VL region that has at least 80% (e.g., 85%, 90%, 95%, 96%,97%, 98%, 99% and any increment therein) sequence identity to a VLregion set forth in SEQ ID NOs: 7, 9, 11, 13, 15, 17, 19, 21, 23, 25,and 27 and a VH region that has at least 80% (e.g., 85%, 90%, 95%, 96%,97%, 98%, 99% and any increment therein) sequence identity to a VHregion set forth in SEQ ID NOs: 8, 10, 12, 14, 16, 18, 20, 22, 24, 26,28, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, and 87. In someembodiments, the anti-Galectin-9 antibody comprises a VL or VH regionthat has at least 80% (e.g., 85%, 90%, 95%, 96%, 97%, 98%, 99% and anyincrement therein) sequence identity to a VL or VH region set forth inSEQ ID NOs: 7-288 and 74-87. In some specific embodiments, theanti-Galectin-9 antibody comprises a VL or VH region that has at least80% (e.g., 85%, 90%, 95%, 96%, 97%, 98%, 99% and any increment therein)sequence identity to a VL or VH region set forth in SEQ ID NO: 7-288 and74-87 and any combination(s) thereof.

In some embodiments, the anti-Galectin-9 antibody comprises a VL regionthat has at least 80% (e.g., 85%, 90%, 95%, 96%, 97%, 98%, 99% and anyincrement therein) sequence identity to a VL region of an antibodyselected from G9.1-1, G9.1-2, G9.1-3, G9.1-4, G9.1-5, G9.1-6, G9.1-7,G9.1-8, G9.1-9, G9.1-10, G9.1-11, G9.1-8m1, G9.1-8m2, G9.1-8m3,G9.1-8m4, G9.1-8m5, G9.1-8m6, G9.1-8m7, G9.1-8m8, G9.1-8m9, G9.1-8m10,G9.1-8m11, G9.1-8m12, G9.1-8m13, and G9.1-8m14. In some embodiments, theanti-Galectin-9 antibody comprises a VH region that has at least 80%(e.g., 85%, 90%, 95%, 96%, 97%, 98%, 99% and any increment therein)sequence identity to a VH region of an antibody selected from G9.1-1,G9.1-2, G9.1-3, G9.1-4, G9.1-5, G9.1-6, G9.1-7, G9.1-8, G9.1-9, G9.1-10,G9.1-11, G9.1-8m1, G9.1-8m2, G9.1-8m3, G9.1-8m4, G9.1-8m5, G9.1-8m6,G9.1-8m7, G9.1-8m8, G9.1-8m9, G9.1-8m10, G9.1-8m11, G9.1-8m12,G9.1-8m13, and G9.1-8m14. In some embodiments, the anti-Galectin-9antibody comprises a VL and a VH region that has at least 80% (e.g.,85%, 90%, 95%, 96%, 97%, 98%, 99% and any increment therein) sequenceidentity to a VL or VH region of an antibody selected from G9.1-1,G9.1-2, G9.1-3, G9.1-4, G9.1-5, G9.1-6, G9.1-7, G9.1-8, G9.1-9, G9.1-10,G9.1-11, G9.1-8m1, G9.1-8m2, G9.1-8m3, G9.1-8m4, G9.1-8m5, G9.1-8m6,G9.1-8m7, G9.1-8m8, G9.1-8m9, G9.1-8m10, G9.1-8m11, G9.1-8m12,G9.1-8m13, and G9.1-8m14.

In some specific embodiments, the anti-Galectin-9 antibody comprises aVL or VH region that has at least 80% (e.g., 85%, 90%, 95%, 96%, 97%,98%, 99% and any increment therein) sequence identity to a VL or VHregion set forth in any of SEQ ID NOs: 21, 22 and 74-87. In somespecific embodiments, the anti-Galectin-9 antibody comprises a VL or VHregion that has at least 80% (e.g., 85%, 90%, 95%, 96%, 97%, 98%, 99%and any increment therein) sequence identity to a VL or VH region setforth in SEQ ID NO: 21, 22 and 74-87 and any combination(s) thereof.

In some specific embodiments, the anti-Galectin-9 antibody comprises aVL region that has at least 80% (e.g., 85%, 90%, 95%, 96%, 97%, 98%, 99%and any increment therein) sequence identity to the VL region ofG9.1-8m13. In some embodiments, the anti-Galectin-9 antibody comprises aVH region that has at least 80% (e.g., 85%, 90%, 95%, 96%, 97%, 98%, 99%and any increment therein) sequence identity to the VH region ofG9.1-8m13. In some embodiments, the anti-Galectin-9 antibody comprises aVL and a VH region that has at least 80% (e.g., 85%, 90%, 95%, 96%, 97%,98%, 99% and any increment therein) sequence identity to the VL or VHregion of G9.1-8m13.

In some embodiments, the anti-Galectin-9 antibody or antigen bindingportion thereof comprises a VL region that has at least 80% (e.g., 85%,90%, 95%, 96%, 97%, 98%, 99% and any increment therein) sequenceidentity to the VL region set forth in SEQ ID NO: 21. In someembodiments, the anti-Galectin-9 antibody or antigen binding portionthereof comprises a VH region that has at least 80% (e.g., 85%, 90%,95%, 96%, 97%, 98%, 99% and any increment therein) sequence identity tothe VH region set forth in SEQ ID NO: 86. In some embodiments, theanti-Galectin-9 antibody or antigen binding portion thereof comprises aVL and VH region that have at least 80% (e.g., 85%, 90%, 95%, 96%, 97%,98%, 99% and any increment therein) sequence identity to the VL and VHregions set forth in SEQ ID NO: 21 and 86.

In some embodiments, the anti-Galectin-9 antibody comprises a VL regionthat has at least 80% (e.g., 85%, 90%, 95%, 96%, 97%, 98%, 99% and anyincrement therein) sequence identity to a VL region set forth in SEQ IDNO: 13, 29, 34, 36, 38, 40, 42, 44, 46, 48, 29, 34, 54, 58, 61, 63, 65,73, 67, 69, and 71. In some embodiments, the anti-Galectin-9 antibodycomprises a VH region that has at least 80% (e.g., 85%, 90%, 95%, 96%,97%, 98%, 99% and any increment therein) sequence identity to a VHregion set forth in SEQ ID NO: 30, 31, 32, 33, 35, 37, 39, 41, 43, 45,47, 49, 50, 51, 52, 53, 55, 56, 57, 59, 60, 62, 64, 66, 68, 70, 72 and73. In some embodiments, the anti-Galectin-9 antibody comprises a VLregion that has at least 80% (e.g., 85%, 90%, 95%, 96%, 97%, 98%, 99%and any increment therein) sequence identity to a VL region set forth inSEQ ID NO: 13, 29, 34, 36, 38, 40, 42, 44, 46, 48, 29, 34, 54, 58, 61,63, 65, 73, 67, 69, and 71 and a VH region that has at least 80% (e.g.,85%, 90%, 95%, 96%, 97%, 98%, 99% and any increment therein) sequenceidentity to a VH region set forth in SEQ ID NO: 30, 31, 32, 33, 35, 37,39, 41, 43, 45, 47, 49, 50, 51, 52, 53, 55, 56, 57, 59, 60, 62, 64, 66,68, 70, 72 and 73. In some specific embodiments, the anti-Galectin-9antibody comprises a VL or VH region that has at least 80% (e.g., 85%,90%, 95%, 96%, 97%, 98%, 99% and any increment therein) sequenceidentity to a VL or VH region set forth in SEQ ID NO: 29-75 and 77-85.In some specific embodiments, the anti-Galectin-9 antibody comprises aVL or VH region that has at least 80% (e.g., 85%, 90%, 95%, 96%, 97%,98%, 99% and any increment therein) sequence identity to a VL or VHregion set forth in SEQ ID NO: 13, 29-73 and any combination(s) thereof.In some specific embodiments, the anti-Galectin-9 antibody comprises aVL or VH region that has at least 80% (e.g., 85%, 90%, 95%, 96%, 97%,98%, 99% and any increment therein) sequence identity to a VL or VHregion set forth in any of SEQ ID NOs: 54, 55, and 56.

In some embodiments, the anti-Galectin-9 antibody comprises a VL regionthat has at least 80% (e.g., 85%, 90%, 95%, 96%, 97%, 98%, 99% and anyincrement therein) sequence identity to a VL region of an antibodyselected from G9.2-1, G9.2-2, G9.2-3, G9.2-4, G9.2-5, G9.2-6, G9.2-7,G9.2-8, G9.2-9, G9.2-10, G9.2-11, G9.2-12, G9.2-13, G9.2-14, G9.2-15,G9.2-16, G9.2-17, G9.2-17mut6, G9.2-18, G9.2-19, G9.2-20, G9.2-21,G9.2-22, G9.2-23, G9.2-24, G9.2-25, G9.2-26, and G9.2-low affinitybinder. In some embodiments, the anti-Galectin-9 antibody comprises a VHregion that has at least 80% (e.g., 85%, 90%, 95%, 96%, 97%, 98%, 99%and any increment therein) sequence identity to a VH region of anantibody selected from G9.2-1, G9.2-2, G9.2-3, G9.2-4, G9.2-5, G9.2-6,G9.2-7, G9.2-8, G9.2-9, G9.2-10, G9.2-11, G9.2-12, G9.2-13, G9.2-14,G9.2-15, G9.2-16, G9.2-17, G9.2-17mut6, G9.2-18, G9.2-19, G9.2-20,G9.2-21, G9.2-22, G9.2-23, G9.2-24, G9.2-25, G9.2-26, and G9.2-lowaffinity binder. In some embodiments, the anti-Galectin-9 antibodycomprises VL and VH regions that has at least 80% (e.g., 85%, 90%, 95%,96%, 97%, 98%, 99% and any increment therein) sequence identity to VLand VH regions of an antibody selected from G9.2-1, G9.2-2, G9.2-3,G9.2-4, G9.2-5, G9.2-6, G9.2-7, G9.2-8, G9.2-9, G9.2-10, G9.2-11,G9.2-12, G9.2-13, G9.2-14, G9.2-15, G9.2-16, G9.2-17, G9.2-17mut6,G9.2-18, G9.2-19, G9.2-20, G9.2-21, G9.2-22, G9.2-23, G9.2-24, G9.2-25,G9.2-26, and G9.2-low affinity binder.

In some embodiments, the anti-Galectin-9 antibody comprises a heavychain CDR having at least 80% (e.g., 80%, 85%, 90%, 95%, 96%, 97%, 98%,99% and any incremental percent therein) sequence identity with asequence selected from any of SEQ ID NO: 8, 10, 12, 14, 16, 18, 20, 22,24, 26, 28, 30, 31, 32, 33, 35, 37, 39, 41, 43, 45, 47, 49, 50, 51, 52,53, 55, 56, 57, 59, 60, 62, 64, 66, 68, 70, 72 73, 74, 75, 76, 77, 78,79, 80, 81, 82, 83, 84, 85, 86, and 87. Alternatively or in addition,the anti-Galectin-9 antibody comprises a light chain CDR having at least80% (e.g., 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% and any incrementalpercent therein) sequence identity with a sequence selected from any ofSEQ ID NO: 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 34, 36, 38, 40,42, 44, 46, 48, 54, 58, 61, 63, 65, 67, 69, 71, and 73.

In some specific embodiments, the anti-Galectin-9 antibody comprises aVL region that has at least 80% (e.g., 85%, 90%, 95%, 96%, 97%, 98%, 99%and any increment therein) sequence identity to the VL region ofG9.2-17. In some specific embodiments, the anti-Galectin-9 antibodycomprises a VH region that has at least 80% (e.g., 85%, 90%, 95%, 96%,97%, 98%, 99% and any increment therein) sequence identity to the VHregion of G9.2-17. In some specific embodiments, the anti-Galectin-9antibody comprises VL and VH regions that has at least 80% (e.g., 85%,90%, 95%, 96%, 97%, 98%, 99% and any increment therein) sequenceidentity to VL and VH regions of G9.2-17.

In some specific embodiments, the anti-Galectin-9 antibody or antigenbinding fragment thereof comprises a VL that has at least 80% (e.g.,85%, 90%, 95%, 96%, 97%, 98%, 99% and any increment therein) sequenceidentity to a VL region set forth in SEQ ID NO: 54. In some specificembodiments, the anti-Galectin-9 antibody or antigen binding fragmentthereof comprises a VH region that has at least 80% (e.g., 85%, 90%,95%, 96%, 97%, 98%, 99% and any increment therein) sequence identity toa VH region set forth in SEQ ID NO: 55. In some specific embodiments,the anti-Galectin-9 antibody or antigen binding fragment thereofcomprises a VL and/or VH region that has at least 80% (e.g., 85%, 90%,95%, 96%, 97%, 98%, 99% and any increment therein) sequence identity toa VL and/or VH region set forth in SEQ ID NO: 54 and 55.

Complementarity Determining Regions (CDRs)

Anti-Galectin-9 antibodies, e.g., binding to CRD1, can comprise thelight and heavy chain CDR1s, CDR2s and CDR3s of G9.1-1, G9.1-2, G9.1-3,G9.1-4, G9.1-5, G9.1-6, G9.1-7, G9.1-8, G9.1-9, G9.1-10, G9.1-11,G9.1-8m1, G9.1-8m2, G9.1-8m3, G9.1-8m4, G9.1-8m5, G9.1-8m6, G9.1-8m7,G9.1-8m8, G9.1-8m9, G9.1-8m10, G9.1-8m11, G9.1-8m12, G9.1-8m13, andG9.1-8m14, or combinations thereof. The amino acid sequence of the VLCDR1s of G9.1-1, G9.1-2, G9.1-3, G9.1-4, G9.1-5, G9.1-6, G9.1-7, G9.1-8,G9.1-9, G9.1-10, G9.1-11, G9.1-8m1, G9.1-8m2, G9.1-8m3, G9.1-8m4,G9.1-8m5, G9.1-8m6, G9.1-8m7, G9.1-8m8, G9.1-8m9, G9.1-8m10, G9.1-8m11,G9.1-8m12, G9.1-8m13, and G9.1-8m14 is set forth in SEQ ID NO: 328. Theamino acid sequence of the VL CDR2s of G9.1-1, G9.1-2, G9.1-3, G9.1-4,G9.1-5, G9.1-6, G9.1-7, G9.1-8, G9.1-9, G9.1-10, G9.1-11, G9.1-8m1,G9.1-8m2, G9.1-8m3, G9.1-8m4, G9.1-8m5, G9.1-8m6, G9.1-8m7, G9.1-8m8,G9.1-8m9, G9.1-8m10, G9.1-8m11, G9.1-8m12, G9.1-8m13, and G9.1-8m14 isset forth in SEQ ID NO: 329. The amino acid sequences of the VL CDR3s ofG9.1-1, G9.1-2, G9.1-3, G9.1-4, G9.1-5, G9.1-6, G9.1-7, G9.1-8, G9.1-9,G9.1-10, G9.1-11, G9.1-8m1, G9.1-8m2, G9.1-8m3, G9.1-8m4, G9.1-8m5,G9.1-8m6, G9.1-8m7, G9.1- 8m8, G9.1-8m9, G9.1-8m10, G9.1-8m11,G9.1-8m12, G9.1-8m13, and G9.1-8m14 are set forth in SEQ ID NO: 330-340.The amino acid sequences of the VH CDR1s of G9.1-1, G9.1-2, G9.1-3,G9.1-4, G9.1-5, G9.1-6, G9.1-7, G9.1-8, G9.1-9, G9.1-10, G9.1-11,G9.1-8m1, G9.1-8m2, G9.1-8m3, G9.1-8m4, G9.1-8m5, G9.1-8m6, G9.1-8m7,G9.1-8m8, G9.1-8m9, G9.1-8m10, G9.1-8m11, G9.1-8m12, G9.1-8m13, andG9.1-8m14 are set forth in SEQ ID NO: 361, 427, 428, 431, 435, 436, 437.The amino acid sequences of the VH CDR2s of G9.1-1, G9.1-2, G9.1-3,G9.1-4, G9.1-5, G9.1-6, G9.1-7, G9.1-8, G9.1-9, G9.1-10, G9.1-11,G9.1-8m1, G9.1-8m2, G9.1-8m3, G9.1-8m4, G9.1-8m5, G9.1-8m6, G9.1-8m7,G9.1-8m8, G9.1-8m9, G9.1-8m10, G9.1-8m11, G9.1-8m12, G9.1-8m13, andG9.1-8m14 are set forth in SEQ ID NO: 362-366, and 438-445. The aminoacid sequences of the VH CDR3s of G9.1-1, G9.1-2, G9.1-3, G9.1-4,G9.1-5, G9.1-6, G9.1-7, G9.1-8, G9.1-9, G9.1-10, G9.1-11, G9.1-8m1,G9.1-8m2, G9.1-8m3, G9.1-8m4, G9.1-8m5, G9.1-8m6, G9.1-8m7, G9.1-8m8,G9.1-8m9, G9.1-8m10, G9.1-8m11, G9.1-8m12, G9.1-8m13, and G9.1-8m14 areset forth in SEQ ID NO: 367-386.

In some embodiments, the anti-Galectin-9 antibody comprises a VL CDR1having the sequence of SEQ ID NO: 328. In some embodiments, theanti-Galectin-9 antibody comprises a VL CDR2 having the sequence of SEQID NO: 329. In some embodiments, the anti-Galectin-9 antibody comprisesa VL CDR3 having a sequence selected from any of SEQ ID NOs: 330-340. Insome embodiments, the anti-Galectin-9 antibody comprises a VL CDR3having the sequence of SEQ ID NO: 337. In some embodiments, theanti-Galectin-9 antibody comprises a VL CDR1 having the sequence of SEQID NO: 328, a VL CDR2 having the sequence of SEQ ID NO: 329, and a VLCDR3 having a sequence selected from any of SEQ ID NOs: 330-340. In someembodiments, the anti-Galectin-9 antibody comprises a VL CDR1 having thesequence of SEQ ID NO: 328, a VL CDR2 having the sequence of SEQ ID NO:329, and a VL CDR3 having the sequence of SEQ ID NO: 337. In someembodiments, the anti-Galectin-9 antibody comprises a VH CDR1 having asequence selected from any of SEQ ID NOs: 361, 427, 428, 431, 435, 436,and 437. In some embodiments, the anti-Galectin-9 antibody comprises aVH CDR1 having the sequence of SEQ ID NO: 361. In some embodiments, theanti-Galectin-9 antibody comprises a VH CDR2 having a sequence selectedfrom any of SEQ ID NOs: 362-366, and 438-445. In some embodiments, theanti-Galectin-9 antibody comprises a VH CDR2 having a sequence selectedfrom SEQ ID NO: 364 or 366. In some embodiments, the anti-Galectin-9antibody comprises a VH CDR3 having a sequence selected from any of SEQID NOs: 367-386. In some embodiments, the anti-Galectin-9 antibodycomprises a VH CDR3 having the sequence of SEQ ID NO: 374 or 383. Insome embodiments, the anti-Galectin-9 antibody comprises a VH CDR1having a sequence selected from any of SEQ ID NOs: 361, 427, 428, 431,435, 436, and 437, a VH CDR2 having a sequence selected from any of SEQID NOs: 362-366 and 438-445, and a VH CDR3 having a sequence selectedfrom any of SEQ ID NOs: 367-386. In some embodiments, theanti-Galectin-9 antibody comprises a VH CDR1 having the sequence of SEQID NO: 361, a VH CDR2 having the sequence of SEQ ID NO: 364, and a VHCDR3 having the sequence of SEQ ID NO: 374. In some embodiments, theanti-Galectin-9 antibody comprises a VH CDR1 having the sequence of SEQID NO: 361, a VH CDR2 having the sequence of SEQ ID NO: 366, and a VHCDR3 having the sequence of SEQ ID NO: 383. In some embodiments, theanti-Galectin-9 antibody comprises a VL CDR1 having the sequence of SEQID NO: 328, a VL CDR2 having the sequence of SEQ ID NO: 329, a VL CDR3having a sequence selected from any of SEQ ID NOs: 330-340, a VH CDR1having a sequence selected from any of SEQ ID NOs: 361, 427, 428, 431,435, 436, and 437, a VH CDR2 having a sequence selected from any of SEQID NOs: 362-366 and 438-445, and a VH CDR3 having a sequence selectedfrom any of SEQ ID NOs: 367-386. In some embodiments, theanti-Galectin-9 antibody comprises a VL CDR1 having the sequence of SEQID NO: 328, a VL CDR2 having the sequence of SEQ ID NO: 329, a VL CDR3having the sequence of SEQ ID NO: 337, a VH CDR1 having the sequence ofSEQ ID NO: 361, a VH CDR2 having the sequence of SEQ ID NO: 364, and aVH CDR3 having the sequence of SEQ ID NO: 374. In some embodiments, theanti-Galectin-9 antibody comprises a VL CDR1 having the sequence of SEQID NO: 328, a VL CDR2 having the sequence of SEQ ID NO: 329, a VL CDR3having the sequence of SEQ ID NO: 337, a VH CDR1 having the sequence ofSEQ ID NO: 361, a VH CDR2 having the sequence of SEQ ID NO: 366, and aVH CDR3 having the sequence of SEQ ID NO: 383. In any of theseembodiments, the anti-Galectin-9 antibody binds to CRD1.

In some embodiments, the anti-Galectin-9 antibodies, e.g., binding toCRD2, comprise the light and heavy chain CDR1s, CDR2s and CDR3s ofG9.2-1, G9.2-2, G9.2-3, G9.2-4, G9.2-5, G9.2-6, G9.2-7, G9.2-8, G9.2-9,G9.2-10, G9.2-11, G9.2-12, G9.2-13, G9.2-14, G9.2-15, G9.2-16, G9.2-17,G9.2-17mut6, G9.2-18, G9.2-19, G9.2-20, G9.2-21, G9.2-22, G9.2-23,G9.2-24, G9.2-25, G9.2-26, and G9.2-low affinity binder, or combinationsthereof. The amino acid sequence of the VL CDR1s of G9.2-1, G9.2-2,G9.2-3, G9.2-4, G9.2-5, G9.2-6, G9.2-7, G9.2-8, G9.2-9, G9.2-10,G9.2-11, G9.2-12, G9.2-13, G9.2-14, G9.2-15, G9.2-16, G9.2-17,G9.2-17mut6, G9.2-18, G9.2-19, G9.2-20, G9.2-21, G9.2-22, G9.2-23,G9.2-24, G9.2-25, G9.2-26, and G9.2-low affinity binder is set forth inSEQ ID NO: 328. The amino acid sequence of the VL CDR2s of G9.2-1,G9.2-2, G9.2-3, G9.2-4, G9.2-5, G9.2-6, G9.2-7, G9.2-8, G9.2-9, G9.2-10,G9.2-11, G9.2-12, G9.2-13, G9.2-14, G9.2-15, G9.2-16, G9.2-17,G9.2-17mut6, G9.2-18, G9.2-19, G9.2-20, G9.2-21, G9.2-22, G9.2-23,G9.2-24, G9.2-25, G9.2-26, and G9.2-low affinity binder is set forth inSEQ ID NO: 329. The amino acid sequences of the VL CDR3s of G9.2-1,G9.2-2, G9.2-3, G9.2-4, G9.2-5, G9.2-6, G9.2-7, G9.2-8, G9.2-9, G9.2-10,G9.2-11, G9.2-12, G9.2-13, G9.2-14, G9.2-15, G9.2-16, G9.2-17,G9.2-17mut6, G9.2-18, G9.2-19, G9.2-20, G9.2-21, G9.2-22, G9.2-23,G9.2-24, G9.2-25, G9.2-26, and G9.2-low affinity binder are set forth inSEQ ID NO: 341-360. The amino acid sequences of the VH CDR1 of G9.2-1,G9.2-2, G9.2-3, G9.2-4, G9.2-5, G9.2-6, G9.2-7, G9.2-8, G9.2-9, G9.2-10,G9.2-11, G9.2-12, G9.2-13, G9.2-14, G9.2-15, G9.2-16, G9.2-17,G9.2-17mut6, G9.2-18, G9.2-19, G9.2-20, G9.2-21, G9.2-22, G9.2-23,G9.2-24, G9.2-25, G9.2-26, and G9.2-low affinity binder are set forth inSEQ ID NO: 361, 424-434. The amino acid sequences of the VH CDR2s ofG9.2-1, G9.2-2, G9.2-3, G9.2-4, G9.2-5, G9.2-6, G9.2-7, G9.2-8, G9.2-9,G9.2-10, G9.2-11, G9.2-12, G9.2-13, G9.2-14, G9.2-15, G9.2-16, G9.2-17,G9.2-17mut6, G9.2-18, G9.2-19, G9.2-20, G9.2-21, G9.2-22, G9.2-23,G9.2-24, G9.2-25, G9.2-26, and G9.2-low affinity binder are set forth inSEQ ID NO: 362, 363, 387-389 and 446-466. The amino acid sequences ofthe VH CDR3s of G9.2-1, G9.2-2, G9.2-3, G9.2-4, G9.2-5, G9.2-6, G9.2-7,G9.2-8, G9.2-9, G9.2-10, G9.2-11, G9.2-12, G9.2-13, G9.2-14, G9.2-15,G9.2-16, G9.2-17, G9.2-17mut6, G9.2-18, G9.2-19, G9.2-20, G9.2-21,G9.2-22, G9.2-23, G9.2-24, G9.2-25, G9.2-26, and G9.2-low affinitybinder are set forth in SEQ ID NO: 390-417.

In some embodiments, the anti-Galectin-9 antibody comprises a VL CDR1having the sequence of SEQ ID NO: 328. In some embodiments, theanti-Galectin-9 antibody comprises a VL CDR2 having the sequence of SEQID NO: 329. In some embodiments, the anti-Galectin-9 antibody comprisesa VL CDR3 having a sequence selected from any of SEQ ID NOs: 341-360. Insome embodiments, the anti-Galectin-9 antibody comprises a VL CDR3having the sequence of SEQ ID NO: 352. In some embodiments, theanti-Galectin-9 antibody comprises a VL CDR1 having the sequence of SEQID NO: 328, a VL CDR2 having the sequence of SEQ ID NO: 329, and a VLCDR3 having a sequence selected from any of SEQ ID NOs: 341-360. In someembodiments, the anti-Galectin-9 antibody comprises a VL CDR1 having thesequence of SEQ ID NO: 328, a VL CDR2 having the sequence of SEQ ID NO:329, and a VL CDR3 having the sequence of SEQ ID NO: 352. In someembodiments, the anti-Galectin-9 antibody comprises a VH CDR1 having asequence selected from any of SEQ ID NOs: 361, and 424-434. In someembodiments, the anti-Galectin-9 antibody comprises a VH CDR1 having thesequence of SEQ ID NO: 361. In some embodiments, the anti-Galectin-9antibody comprises a VH CDR2 having a sequence selected from any of SEQID NOs: 362, 363, 387-389 and 446-466. In some embodiments, theanti-Galectin-9 antibody comprises a VH CDR2 having the sequence of SEQID NO: 388. In some embodiments, the anti-Galectin-9 antibody comprisesa VH CDR3 having a sequence selected from any of SEQ ID NOs: 390-417. Insome embodiments, the anti-Galectin-9 antibody comprises a VH CDR3having the sequence of SEQ ID NO: 406 or 407. In some embodiments, theanti-Galectin-9 antibody comprises a VH CDR1 having a sequence selectedfrom any of SEQ ID NOs: 361, and 424-434, a VH CDR2 having a sequenceselected from any of SEQ ID NOs: 362, 363, 387-389 and 446-466, and a VHCDR3 having a sequence selected from any of SEQ ID NOs: 390-417. In someembodiments, the anti-Galectin-9 antibody comprises a VH CDR1 having thesequence of SEQ ID NO: 361, a VH CDR2 having the sequence of SEQ ID NO:388, and a VH CDR3 having the sequence of SEQ ID NO: 406. In someembodiments, the anti-Galectin-9 antibody comprises a VH CDR1 having thesequence of SEQ ID NO: 361, a VH CDR2 having the sequence of SEQ ID NO:388, and a VH CDR3 having the sequence of SEQ ID NO: 407. In someembodiments, the anti-Galectin-9 antibody comprises a VL CDR1 having thesequence of SEQ ID NO: 328, a VL CDR2 having the sequence of SEQ ID NO:329, a VL CDR3 having a sequence selected from any of SEQ ID NOs:341-360, a VH CDR1 having a sequence selected from any of SEQ ID NOs:361, and 424-434, a VH CDR2 having a sequence selected from any of SEQID NOs: 362, 363, 387-389 and 446-466, and a VH CDR3 having a sequenceselected from any of SEQ ID NOs: 390-417. In some embodiments, theanti-Galectin-9 antibody comprises a VL CDR1 having the sequence of SEQID NO: 328, a VL CDR2 having the sequence of SEQ ID NO: 329, a VL CDR3having the sequence of SEQ ID NO: 352, a VH CDR1 having the sequence ofSEQ ID NO: 361, a VH CDR2 having the sequence of SEQ ID NO: 388, and aVH CDR3 having the sequence of SEQ ID NO: 406. In some embodiments, theanti-Galectin-9 antibody comprises a VL CDR1 having the sequence of SEQID NO: 328, a VL CDR2 having the sequence of SEQ ID NO: 329, a VL CDR3having the sequence of SEQ ID NO: 352, a VH CDR1 having the sequence ofSEQ ID NO: 361, a VH CDR2 having the sequence of SEQ ID NO: 388, and aVH CDR3 having the sequence of SEQ ID NO: 407. In any of theseembodiments, the anti-Galectin-9 antibody binds to CRD1.

Because Galectin-9 binding specificity is dictated essentially by theCDR1, 2 and 3 regions, the VH CDR1, 2 and 3 sequences and the VL CDR1, 2and 3 sequences disclosed above, can be mixed and matched to generatenew Galectin-9 binding antibodies, as long as each resulting newantibody has a VL CDR1, 2 and 3 and a VH CDR1, 2 and 3. Such antibodiesresulting from a new combination of CDRs described herein can be testedusing the binding assays described herein. In some embodiments, theCDR1, CDR2 and/or CDR3 sequence from a particular VH or VL sequence isreplaced with a structurally similar CDR sequence(s). Novel VH and VLsequences can be created by substituting one or more VH and/or VL CDRsequence(s) with structurally similar sequences from the CDR sequencesdisclosed herein, according to methods known in the art.

Accordingly, in some embodiments, anti-Galectin-9 antibodies or antigenbinding portions thereof comprise (a) VL CDR1 amino acid sequence setforth in SEQ ID NO: 328; (b) VL CDR2 amino acid sequence set forth inSEQ ID NO: 329; (c) VL CDR3 amino acid sequence selected from SEQ ID NO:330-340 and 341-360 (d) VH CDR1 amino acid sequence set forth in SEQ IDNO: SEQ ID NO: 361, 427, 428, 431, 435, 436, 437; and SEQ ID NO: 361,424-434 (e) VH CDR2 amino acid sequence selected from SEQ ID NO: 362-366and 438-445, 362, 363, and 387-389 and 446-466; (f) VH CDR3 amino acidsequence selected from SEQ ID NO: 367-386 and 390-417.

In some embodiments, anti-Galectin-9 antibodies or antigen bindingportions thereof comprise a VL CDR1 amino acid sequence set forth in SEQID NO: 328. In some embodiments, anti-Galectin-9 antibodies or antigenbinding portions thereof comprise a VL CDR2 amino acid sequence setforth in SEQ ID NO: 329. In some embodiments, anti-Galectin-9 antibodiesor antigen binding portions thereof comprise a VL CDR3 amino acidsequence selected from SEQ ID NO: 330-340. In some embodiments,anti-Galectin-9 antibodies or antigen binding portions thereof comprisea VH CDR1 amino acid sequence set forth in SEQ ID NO: 361, 427, 428,431, 435, 436, 437. In some embodiments, anti-Galectin-9 antibodies orantigen binding portions thereof comprise a VH CDR2 amino acid sequenceselected from SEQ ID NO: 362-366, and 438-445. In some embodiments,anti-Galectin-9 antibodies or antigen binding portions thereof comprisea VH CDR3 amino acid sequence selected from SEQ ID NO: 367-386.Accordingly, in some embodiments, anti-Galectin-9 antibodies or antigenbinding portions thereof comprise (a) VL CDR1 amino acid sequence setforth in SEQ ID NO: 328; (b) VL CDR2 amino acid sequence set forth inSEQ ID NO: 329; (c) VL CDR3 amino acid sequence selected from SEQ ID NO:330-340; (d) VH CDR1 amino acid sequence set forth in SEQ ID NO: 361,427, 428, 431, 435, 436, 437; (e) VH CDR2 amino acid sequence selectedfrom SEQ ID NO: 362-366 and 438-445; (f) VH CDR3 amino acid sequenceselected from SEQ ID NO: 367-386.

In some embodiments, anti-Galectin-9 antibodies or antigen bindingportions thereof comprise a VL CDR1 amino acid sequence set forth in SEQID NO: 328. In some embodiments, anti-Galectin-9 antibodies or antigenbinding portions thereof comprise a VL CDR2 amino acid sequence setforth in SEQ ID NO: 329. In some embodiments, anti-Galectin-9 antibodiesor antigen binding portions thereof comprise a VL CDR3 amino acidsequence selected from SEQ ID NO: 341-360. In some embodiments,anti-Galectin-9 antibodies or antigen binding portions thereof comprisea VH CDR1 amino acid sequence set forth in SEQ ID NO: 361, 424-434. Insome embodiments, anti-Galectin-9 antibodies or antigen binding portionsthereof comprise a VH CDR2 amino acid sequence selected from SEQ ID NO:362, 363, 387-389 and 446-466. In some embodiments, anti-Galectin-9antibodies or antigen binding portions thereof comprise a VH CDR3 aminoacid sequence selected from SEQ ID NO: 390-417.

Accordingly, in some embodiments, anti-Galectin-9 antibodies or antigenbinding portions thereof comprise (a) VL CDR1 amino acid sequence setforth in SEQ ID NO: 328; (b) VL CDR2 amino acid sequence set forth inSEQ ID NO: 329; (c) VL CDR3 amino acid sequence selected from SEQ ID NO:341-360; (d) VH CDR1 amino acid sequence set forth in SEQ ID NO: 361;(e) VH CDR2s amino acid sequence selected from SEQ ID NO: 362, 363,387-389 and 446-466; (f) VH CDR3 amino acid sequence selected from SEQID NO: 390-417.

9.1 Antibody Clones and Related CDRs

Clone 9.1-Derived Light Chain Variable Regions

In some embodiments, the anti-Galectin-9 antibody or antigen bindingportion thereof comprises heavy and light chain variable regions,wherein the light chain variable region CDR1, CDR2, and CDR3 compriseSEQ ID NO: 328, 329, and 330, respectively. In some embodiments, thelight chain variable region CDR1, CDR2, and CDR3 consist of SEQ ID NO:328, 329, and 330, respectively. In some embodiments, the antibodycomprises the same VL CDRs as G9.1-1.

In some embodiments, the anti-Galectin-9 antibody or antigen bindingportion thereof comprises heavy and light chain variable regions,wherein the light chain variable region CDR1, CDR2, and CDR3 compriseSEQ ID NO: 328, 329, and 331, respectively. In some embodiments, thelight chain variable region CDR1, CDR2, and CDR3 consist of SEQ ID NO:328, 329, and 331, respectively. In some embodiments, the antibodycomprises the same VL CDRs as G9.1-2.

In some embodiments, the anti-Galectin-9 antibody or antigen bindingportion thereof comprises heavy and light chain variable regions,wherein the light chain variable region CDR1, CDR2, and CDR3 compriseSEQ ID NO: 328, 329, and 332, respectively. In some embodiments, thelight chain variable region CDR1, CDR2, and CDR3 consist of SEQ ID NO:328, 329, and 332, respectively. In some embodiments, the antibodycomprises the same VL CDRs as G9.1-3.

In some embodiments, the anti-Galectin-9 antibody or antigen bindingportion thereof comprises heavy and light chain variable regions,wherein the light chain variable region CDR1, CDR2, and CDR3 compriseSEQ ID NO: 328, 329, and 333, respectively. In some embodiments, thelight chain variable region CDR1, CDR2, and CDR3 consist of SEQ ID NO:328, 329, and 333, respectively. In some embodiments, the antibodycomprises the same VL CDRs as G9.1-4.

n some embodiments, the anti-Galectin-9 antibody or antigen bindingportion thereof comprises heavy and light chain variable regions,wherein the light chain variable region CDR1, CDR2, and CDR3 compriseSEQ ID NO: 328, 329, and 334, respectively. In some embodiments, thelight chain variable region CDR1, CDR2, and CDR3 consist of SEQ ID NO:328, 329, and 334, respectively. In some embodiments, the antibodycomprises the same VL CDRs as G9.1-5.

In some embodiments, the anti-Galectin-9 antibody or antigen bindingportion thereof comprises heavy and light chain variable regions,wherein the light chain variable region CDR1, CDR2, and CDR3 compriseSEQ ID NO: 328, 329, and 335, respectively. In some embodiments, thelight chain variable region CDR1, CDR2, and CDR3 consist of SEQ ID NO:328, 329, and 335, respectively. In some embodiments, the antibodycomprises the same VL CDRs as G9.1-6.

n some embodiments, the anti-Galectin-9 antibody or antigen bindingportion thereof comprises heavy and light chain variable regions,wherein the light chain variable region CDR1, CDR2, and CDR3 compriseSEQ ID NO: 328, 329, and 336, respectively. In some embodiments, thelight chain variable region CDR1, CDR2, and CDR3 consist of SEQ ID NO:328, 329, and 336, respectively. In some embodiments, the antibodycomprises the same VL CDRs as G9.1-7.

In some embodiments, the anti-Galectin-9 antibody or antigen bindingportion thereof comprises heavy and light chain variable regions,wherein the light chain variable region CDR1, CDR2, and CDR3 compriseSEQ ID NO: 328, 329, and 337, respectively. In some embodiments, thelight chain variable region CDR1, CDR2, and CDR3 consist of SEQ ID NO:328, 329, and 337, respectively. In some embodiments, the antibodycomprises the same VL CDRs as G9.1-8.

In some embodiments, the anti-Galectin-9 antibody or antigen bindingportion thereof comprises heavy and light chain variable regions,wherein the light chain variable region CDR1, CDR2, and CDR3 compriseSEQ ID NO: 328, 329, and 338, respectively. In some embodiments, thelight chain variable region CDR1, CDR2, and CDR3 consist of SEQ ID NO:328, 329, and 338, respectively. In some embodiments, the antibodycomprises the same VL CDRs as G9.1-9.

In some embodiments, the anti-Galectin-9 antibody or antigen bindingportion thereof comprises heavy and light chain variable regions,wherein the light chain variable region CDR1, CDR2, and CDR3 compriseSEQ ID NO: 328, 329, and 339, respectively. In some embodiments, thelight chain variable region CDR1, CDR2, and CDR3 consist of SEQ ID NO:328, 329, and 339, respectively. In some embodiments, the antibodycomprises the same VL CDRs as G9.1-10.

In some embodiments, the anti-Galectin-9 antibody or antigen bindingportion thereof comprises heavy and light chain variable regions,wherein the light chain variable region CDR1, CDR2, and CDR3 compriseSEQ ID NO: 328, 329, and 340, respectively. In some embodiments, thelight chain variable region CDR1, CDR2, and CDR3 consist of SEQ ID NO:328, 329, and 340, respectively. In some embodiments, the antibodycomprises the same VL CDRs as G9.1-11.

In some embodiments, the anti-Galectin-9 antibody or antigen bindingportion thereof comprises heavy and light chain variable regions,wherein the light chain variable region CDR1 comprises SEQ ID NO: 328.In some embodiments, the anti-Galectin-9 antibody or antigen bindingportion thereof comprises heavy and light chain variable regions,wherein the light chain variable region CDR2 comprises SEQ ID NO: 329.In some embodiments, the anti-Galectin-9 antibody or antigen bindingportion thereof comprises heavy and light chain variable regions,wherein the light chain variable region CDR3 comprises SEQ ID NO: 337.In some embodiments, the anti-Galectin-9 antibody or antigen bindingportion thereof comprises heavy and light chain variable regions,wherein the light chain variable region CDR1 consists of SEQ ID NO: 328.In some embodiments, the anti-Galectin-9 antibody or antigen bindingportion thereof comprises heavy and light chain variable regions,wherein the light chain variable region CDR2 consists of SEQ ID NO: 329.In some embodiments, the anti-Galectin-9 antibody or antigen bindingportion thereof comprises heavy and light chain variable regions,wherein the light chain variable region CDR3 consists of SEQ ID NO: 337.In some embodiments, the antibody comprises the same VL CDRs as 9.1-8,G9.1-8m1, G9.1-8m2, G9.1-8m3, G9.1-8m4, G9.1-8m5, G9.1-8m6, G9.1-8m7,G9.1-8m8, G9.1-8m9, G9.1-8m10, G9.1-8m11, G9.1-8m12, G9.1-8m13, orG9.1-8m14. In some embodiments, the antibody comprises the same VL CDRsas G9.1-8m12. In some embodiments, the antibody comprises the same VLCDRs as G9.1-8m13.

In some embodiments, the anti-Galectin-9 antibody or binding portionthereof comprises heavy and light chain variable regions, wherein theheavy chain variable region CDR1 comprises SEQ ID NO: 328. In someembodiments, the anti-Galectin-9 antibody or binding portion thereofcomprises heavy and light chain variable regions, wherein the lightchain variable region CDR2 comprises X₁X₂X₃X₄X₅SX₆X₇X₈SYADSVKG (SEQ IDNO: 467), in which X₁═Y or S, X₂═I or S, X₃═Y or S, X₄═P or S, X₅═Y orS, X₆=G or S, X₇═Y or S, and X₈=T or S. In some embodiments, theanti-Galectin-9 antibody or binding portion thereof comprises heavy andlight chain variable regions, wherein the light chain variable regionCDR3 comprises X₁SX₂X₃X₄X₅X₆X₇X₈X₉X₁₀KX₁₁X₁₂X₁₃GMDY (SEQ ID NO: 468), inwhich X₁═Y or S, X₂=T, S, or absent, X₃═Y, S, or absent, X₄═S or absent,X₅═W, S, or absent, X₆═S or absent, X₇=G, S, or absent, X₈=G, T, S, orabsent, X₉═I, Y, S, or absent, X₁₀=G, S, or Y, X₁₁═W or S, X₁₂═V or S,and X₁₃═W or S. In some examples, the anti-Galectin-9 antibody containsG at X₇, Y at X₈, and/or T at X₉ in the heavy chain CDR2 domain.Alternatively, or in addition, the anti-Galectin-9 antibody containsdeletions at one or more of X₄-X₇ in the heavy chain CDR3 domain. Inother examples, the anti-Galectin-9 antibody contains S at one or moreof X₆-X₈ in the heavy chain CDR2 domain. Alternatively or in addition,the anti-Galectin-9 antibody contains deletions at one or more of X₅-X₇in the heavy chain CDR3 domain. In a further example, theanti-Galectin-9 antibody contains S at one or more of X₆-X₈ in the heavychain CDR2 domain. Alternatively or in addition, the anti-Galectin-9antibody contains deletions at one or more of X₃-X₉ and/or X₁₀ is Yinthe heavy chain CDR3 domain.

Clone 9.1-Derived Heavy Chain Variable Regions

In some embodiments, the anti-Galectin-9 antibody or antigen bindingportion thereof comprises heavy and light chain variable regions,wherein the heavy chain variable region CDR1, CDR2, and CDR3 compriseSEQ ID NO: 431, 438, and 367, respectively. In some embodiments, theheavy chain variable region CDR1, CDR2, and CDR3 consist of SEQ ID NO:431, 438, and 367, respectively. In some embodiments, the antibodycomprises the same VH CDRs as G9.1-1.

In some embodiments, the anti-Galectin-9 antibody or antigen bindingportion thereof comprises heavy and light chain variable regions,wherein the heavy chain variable region CDR1, CDR2, and CDR3 compriseSEQ ID NO: 435, 439, and 368, respectively. In some embodiments, theheavy chain variable region CDR1, CDR2, and CDR3 consist of SEQ ID NO:435, 439, and 368, respectively. In some embodiments, the antibodycomprises the same VH CDRs as G9.1-2.

In some embodiments, the anti-Galectin-9 antibody or antigen bindingportion thereof comprises heavy and light chain variable regions,wherein the heavy chain variable region CDR1, CDR2, and CDR3 compriseSEQ ID NO: 436, 363, and 369, respectively. In some embodiments, theheavy chain variable region CDR1, CDR2, and CDR3 consist of SEQ ID NO:436, 363, and 369, respectively. In some embodiments, the antibodycomprises the same VH CDRs as G9.1-3.

In some embodiments, the anti-Galectin-9 antibody or antigen bindingportion thereof comprises heavy and light chain variable regions,wherein the heavy chain variable region CDR1, CDR2, and CDR3 compriseSEQ ID NO: 437, 440, and 370, respectively. In some embodiments, theheavy chain variable region CDR1, CDR2, and CDR3 consist of SEQ ID NO:437, 440, and 370, respectively. In some embodiments, the antibodycomprises the same VH CDRs as G9.1-4.

In some embodiments, the anti-Galectin-9 antibody or antigen bindingportion thereof comprises heavy and light chain variable regions,wherein the heavy chain variable region CDR1, CDR2, and CDR3 compriseSEQ ID NO: 437, 441, and 371, respectively. In some embodiments, theheavy chain variable region CDR1, CDR2, and CDR3 consist of SEQ ID NO:437, 441, and 371, respectively. In some embodiments, the antibodycomprises the same VH CDRs as G9.1-5.

In some embodiments, the anti-Galectin-9 antibody or antigen bindingportion thereof comprises heavy and light chain variable regions,wherein the heavy chain variable region CDR1, CDR2, and CDR3 compriseSEQ ID NO: 427, 442, and 372, respectively. In some embodiments, theheavy chain variable region CDR1, CDR2, and CDR3 consist of SEQ ID NO:427, 442, and 372, respectively. In some embodiments, the antibodycomprises the same VH CDRs as G9.1-6.

In some embodiments, the anti-Galectin-9 antibody or antigen bindingportion thereof comprises heavy and light chain variable regions,wherein the heavy chain variable region CDR1, CDR2, and CDR3 compriseSEQ ID NO: 361, 443, and 373, respectively. In some embodiments, theheavy chain variable region CDR1, CDR2, and CDR3 consist of SEQ ID NO:361, 443, and 373, respectively. In some embodiments, the antibodycomprises the same VH CDRs as G9.1-7.

In some embodiments, the anti-Galectin-9 antibody or antigen bindingportion thereof comprises heavy and light chain variable regions,wherein the heavy chain variable region CDR1, CDR2, and CDR3 compriseSEQ ID NO: 361, 364, and 374, respectively. In some embodiments, theheavy chain variable region CDR1, CDR2, and CDR3 consist of SEQ ID NO:361, 364, and 374, respectively. In some embodiments, the antibodycomprises the same VH CDRs as G9.1-8.

In some embodiments, the anti-Galectin-9 antibody or antigen bindingportion thereof comprises heavy and light chain variable regions,wherein the heavy chain variable region CDR1, CDR2, and CDR3 compriseSEQ ID NO: 361, 363, and 384, respectively. In some embodiments, theheavy chain variable region CDR1, CDR2, and CDR3 consist of SEQ ID NO:361, 363, and 384, respectively. In some embodiments, the antibodycomprises the same VH CDRs as G9.1-9.

In some embodiments, the anti-Galectin-9 antibody or binding portionthereof comprises heavy and light chain variable regions, wherein theheavy chain variable region CDR1, CDR2, and CDR3 comprise SEQ ID NO:429, 444, and 385, respectively. In some embodiments, the heavy chainvariable region CDR1, CDR2, and CDR3 consist of SEQ ID NO: 429, 444, and385, respectively. In some embodiments, the antibody comprises the sameVH CDRs as G9.1-10. In some embodiments, the anti-Galectin-9 antibody orantigen binding portion thereof comprises heavy and light chain variableregions, wherein the heavy chain variable region CDR1, CDR2, and CDR3comprise SEQ ID NO: 428, 445, and 386, respectively. In someembodiments, the heavy chain variable region CDR1, CDR2, and CDR3consist of SEQ ID NO: 361, 445, and 386, respectively. In someembodiments, the antibody comprises the same VH CDRs as G9.1-11.

In some embodiments, the anti-Galectin-9 antibody or antigen bindingportion thereof comprises heavy and light chain variable regions,wherein the heavy chain variable region CDR1, CDR2, and CDR3 compriseSEQ ID NO: 361, 365, and 374, respectively. In some embodiments, theheavy chain variable region CDR1, CDR2, and CDR3 consist of SEQ ID NO:361, 365, and 374, respectively. In some embodiments, the antibodycomprises the same VH CDRs as G9.1-8m1.

In some embodiments, the anti-Galectin-9 antibody or antigen bindingportion thereof comprises heavy and light chain variable regions,wherein the heavy chain variable region CDR1, CDR2, and CDR3 compriseSEQ ID NO: 361, 366, and 374, respectively. In some embodiments, theheavy chain variable region CDR1, CDR2, and CDR3 consist of SEQ ID NO:361, 366, and 374, respectively. In some embodiments, the antibodycomprises the same VH CDRs as G9.1-8m2.

In some embodiments, the anti-Galectin-9 antibody or antigen bindingportion thereof comprises heavy and light chain variable regions,wherein the heavy chain variable region CDR1, CDR2, and CDR3 compriseSEQ ID NO: 361, 364, and 375, respectively. In some embodiments, theheavy chain variable region CDR1, CDR2, and CDR3 consist of SEQ ID NO:361, 364, and 375, respectively. In some embodiments, the antibodycomprises the same VH CDRs as G9.1-8m3.

In some embodiments, the anti-Galectin-9 antibody or antigen bindingportion thereof comprises heavy and light chain variable regions,wherein the heavy chain variable region CDR1, CDR2, and CDR3 compriseSEQ ID NO: 361, 364, and 376, respectively. In some embodiments, theheavy chain variable region CDR1, CDR2, and CDR3 regions consist of SEQID NO: 361, 364, and 376, respectively. In some embodiments, theantibody comprises the same VH CDRs as G9.1-8m4.

In some embodiments, the anti-Galectin-9 antibody or antigen bindingportion thereof comprises heavy and light chain variable regions,wherein the heavy chain variable region CDR1, CDR2, and CDR3 compriseSEQ ID NO: 361, 364, and 377, respectively. In some embodiments, theheavy chain variable region CDR1, CDR2, and CDR3 consist of SEQ ID NO:361, 364, and 377, respectively. In some embodiments, the antibodycomprises the same VH CDRs as G9.1-8m5.

In some embodiments, the anti-Galectin-9 antibody or antigen bindingportion thereof comprises heavy and light chain variable regions,wherein the heavy chain variable region CDR1, CDR2, and CDR3 compriseSEQ ID NO: 361, 364, and 378, respectively. In some embodiments, theheavy chain variable region CDR1, CDR2, and CDR3 consist of SEQ ID NO:361, 364, and 378, respectively. In some embodiments, the antibodycomprises the same VH CDRs as G9.1-8m6.

In some embodiments, the anti-Galectin-9 antibody or antigen bindingportion thereof comprises heavy and light chain variable regions,wherein the heavy chain variable region CDR1, CDR2, and CDR3 compriseSEQ ID NO: 361, 364, and 379, respectively. In some embodiments, theheavy chain variable region CDR1, CDR2, and CDR3 consist of SEQ ID NO:361, 364, and 379, respectively. In some embodiments, the antibodycomprises the same VH CDRs as G9.1-8m7.

In some embodiments, the anti-Galectin-9 antibody or antigen bindingportion thereof comprises heavy and light chain variable regions,wherein the heavy chain variable region CDR1, CDR2, and CDR3 compriseSEQ ID NO: 361, 364, and 380, respectively. In some embodiments, theheavy chain variable region CDR1, CDR2, and CDR3 consist of SEQ ID NO:361, 364, and 380, respectively. In some embodiments, the antibodycomprises the same VH CDRs as G9.1-8m8.

In some embodiments, the anti-Galectin-9 antibody or antigen bindingportion thereof comprises heavy and light chain variable regions,wherein the heavy chain variable region CDR1, CDR2, and CDR3 compriseSEQ ID NO: 361, 364, and 383, respectively. In some embodiments, theheavy chain variable region CDR1, CDR2, and CDR3 consist of SEQ ID NO:361, 364, and 383, respectively. In some embodiments, the antibodycomprises the same VH CDRs as G9.1-8m9.

In some embodiments, the anti-Galectin-9 antibody or antigen bindingportion thereof comprises heavy and light chain variable regions,wherein the heavy chain variable region CDR1, CDR2, and CDR3 compriseSEQ ID NO: 361, 364, and 381, respectively. In some embodiments, theheavy chain variable region CDR1, CDR2, and CDR3 consist of SEQ ID NO:361, 364, and 381, respectively. In some embodiments, the antibodycomprises the same VH CDRs as G9.1-8m10.

In some embodiments, the anti-Galectin-9 antibody or antigen bindingportion thereof comprises heavy and light chain variable regions,wherein the heavy chain variable region CDR1, CDR2, and CDR3 compriseSEQ ID NO: 361, 364, and 382, respectively. In some embodiments, theheavy chain variable region CDR1, CDR2, and CDR3 consist of SEQ ID NO:361, 364, and 382, respectively. In some embodiments, the antibodycomprises the same VH CDRs as G9.1-8m11.

In some embodiments, the anti-Galectin-9 antibody or antigen bindingportion thereof comprises heavy and light chain variable regions,wherein the heavy chain variable region CDR1, CDR2, and CDR3 compriseSEQ ID NO: 361, 366, and 380, respectively. In some embodiments, theheavy chain variable region CDR1, CDR2, and CDR3 consist of SEQ ID NO:361, 366, and 380, respectively. In some embodiments, the antibodycomprises the same VH CDRs as G9.1-8m12.

In some embodiments, the anti-Galectin-9 antibody or antigen bindingportion thereof comprises heavy and light chain variable regions,wherein the heavy chain variable region CDR1 comprises SEQ ID NO: 361.In some embodiments, the anti-Galectin-9 antibody or antigen bindingportion thereof comprises heavy and light chain variable regions,wherein the heavy chain variable region CDR2 comprises SEQ ID NO: 366.In some embodiments, the anti-Galectin-9 antibody or antigen bindingportion thereof comprises heavy and light chain variable regions,wherein the heavy chain variable region CDR3 region comprises SEQ ID NO:383. In some embodiments, the anti-Galectin-9 antibody or antigenbinding portion thereof comprises heavy and light chain variableregions, wherein the heavy chain variable region CDR1 consists of SEQ IDNO: 361. In some embodiments, the anti-Galectin-9 antibody or antigenbinding portion thereof comprises heavy and light chain variableregions, wherein the heavy chain variable region CDR2 consists of SEQ IDNO: 366. In some embodiments, the anti-Galectin-9 antibody or antigenbinding portion thereof comprises heavy and light chain variableregions, wherein the heavy chain variable region CDR3 region consists ofSEQ ID NO: 383. In some embodiments, the anti-Galectin-9 antibody orantigen binding portion thereof comprises heavy and light chain variableregions, wherein the heavy chain variable region CDR1, CDR2, and CDR3comprise SEQ ID NO: 361, 366, and 383, respectively. In someembodiments, the heavy chain variable region CDR1, CDR2, and CDR3consist of SEQ ID NO: 361, 366, and 383, respectively. In someembodiments, the antibody comprises the same VH CDRs as G9.1-8m13.

In some embodiments, the anti-Galectin-9 antibody or antigen bindingportion thereof comprises heavy and light chain variable regions,wherein the heavy chain variable region CDR1, CDR2, and CDR3 compriseSEQ ID NO: 361, 366, and 382, respectively. In some embodiments, theheavy chain variable region CDR1, CDR2, and CDR3 consist of SEQ ID NO:361, 366, and 382, respectively. In some embodiments, the antibodycomprises the same VH CDRs as G9.1-8m14.

Clone 9.1-9.1 Heavy and Light Chain Variable Regions

In one specific embodiment, the anti-Galectin-9 antibody or antigenbinding portion thereof comprises heavy and light chain variableregions, wherein: the light chain variable region CDR1, CDR2, and CDR3comprise SEQ ID NO: 328, 329, and 330, respectively, and the heavy chainvariable region CDR1, CDR2, and CDR3 comprise SEQ ID NO: 431, 438, and367, respectively. In some embodiments, the light and heavy chainvariable region CDR1, CDR2, and CDR3 consist of SEQ ID NO: 328, 329, and330 and SEQ ID NO: 431, 438, and 367, respectively. In some embodiments,the antibody comprises the same VL and VH CDRs as G9.1-1.

In one specific embodiment, the anti-Galectin-9 antibody or bindingportion thereof comprises heavy and light chain variable regions,wherein: the light chain variable region CDR1, CDR2, and CDR3 compriseSEQ ID NO: 328, 329, and 331, respectively, and the heavy chain variableregion CDR1, CDR2, and CDR3 comprise SEQ ID NO: 435, 439, and 368,respectively. In some embodiments, the light and heavy chain variableregion CDR1, CDR2, and CDR3 regions consist of SEQ ID NO: 328, 329, and331 and SEQ ID NO: 435, 439, and 368. In one specific embodiment, theantibody comprises the same VL and VH CDRs as G9.1-2. In one specificembodiment, the anti-Galectin-9 antibody or binding portion thereofcomprises heavy and light chain variable regions, wherein: the lightchain variable region CDR1, CDR2, and CDR3 comprise SEQ ID NO: 328, 329,and 332, respectively, and the heavy chain variable region CDR1, CDR2,and CDR3 comprise SEQ ID NO: 436, 363, and 369, respectively. In someembodiments, the light and heavy chain variable region CDR1, CDR2, andCDR3 regions consist of SEQ ID NO: 328, 329, and 332 and SEQ ID NO: 436,363, and 369. In one specific embodiment, the antibody comprises thesame VL and VH CDRs as G9.1-3.

In one specific embodiment, the anti-Galectin-9 antibody or bindingportion thereof comprises heavy and light chain variable regions,wherein: the light chain variable region CDR1, CDR2, and CDR3 compriseSEQ ID NO: 328, 329, and 333, respectively, and the heavy chain variableregion CDR1, CDR2, and CDR3 comprise SEQ ID NO: 437, 440, and 370,respectively. In some embodiments, the light and heavy chain variableregion CDR1, CDR2, and CDR3 regions consist of SEQ ID NO: 328, 329, and333, and SEQ ID NO: 437, 440, and 370. In one specific embodiment, theantibody comprises the same VL and VH CDRs as G9.1-4.

In one specific embodiment, the anti-Galectin-9 antibody or bindingportion thereof comprises heavy and light chain variable regions,wherein: the light chain variable region CDR1, CDR2, and CDR3 compriseSEQ ID NO: 328, 329, and 334, respectively, and the heavy chain variableregion CDR1, CDR2, and CDR3 comprise SEQ ID NO: 437, 441, and 371,respectively. In some embodiments, the light and heavy chain variableregion CDR1, CDR2, and CDR3 regions consist of SEQ ID NO: 328, 329, and334, and SEQ ID NO: 437, 441, and 371. In one specific embodiment, theantibody comprises the same VL and VH CDRs as G9.1-5.

In one specific embodiment, the anti-Galectin-9 antibody or bindingportion thereof comprises heavy and light chain variable regions,wherein: the light chain variable region CDR1, CDR2, and CDR3 compriseSEQ ID NO: 328, 329, and 335, respectively, and the heavy chain variableregion CDR1, CDR2, and CDR3 comprise SEQ ID NO: 427, 442, and 372,respectively. In some embodiments, the light and heavy chain variableregion CDR1, CDR2, and CDR3 regions consist of SEQ ID NO: 328, 329, and335, and SEQ ID NO: 427, 442, and 372. In one specific embodiment, theantibody comprises the same VL and VH CDRs as G9.1-6.

In one specific embodiment, the anti-Galectin-9 antibody or bindingportion thereof comprises heavy and light chain variable regions,wherein: the light chain variable region CDR1, CDR2, and CDR3 compriseSEQ ID NO: 328, 329, and 336, respectively, and the heavy chain variableregion CDR1, CDR2, and CDR3 comprise SEQ ID NO: 361, 443, and 373,respectively. In some embodiments, the light and heavy chain variableregion CDR1, CDR2, and CDR3 regions consist of SEQ ID NO: 328, 329, and336, and SEQ ID NO: 361, 443, and 373. In one specific embodiment, theantibody comprises the same VL and VH CDRs as G9.1-7.

In one specific embodiment, the anti-Galectin-9 antibody or bindingportion thereof comprises heavy and light chain variable regions,wherein: the light chain variable region CDR1, CDR2, and CDR3 compriseSEQ ID NO: 328, 329, and 337, respectively, and the heavy chain variableregion CDR1, CDR2, and CDR3 comprise SEQ ID NO: 361, 364, and 374. Insome embodiments, the light and heavy chain variable region CDR1, CDR2,and CDR3 regions consist of SEQ ID NO: 328, 329, and 337, and SEQ ID NO:361, 364, and 374. In one specific embodiment, the antibody comprisesthe same VL and VH CDRs as G9.1-8.

In one specific embodiment, the anti-Galectin-9 antibody or bindingportion thereof comprises heavy and light chain variable regions,wherein: the light chain variable region CDR1, CDR2, and CDR3 compriseSEQ ID NO: 328, 329, and 338, respectively, and the heavy chain variableregion CDR1, CDR2, and CDR3 comprise SEQ ID NO: 361, 363, and 384. Insome embodiments, the light and heavy chain variable region CDR1, CDR2,and CDR3 regions consist of SEQ ID NO: 328, 329, and 338, and SEQ ID NO:361, 363, and 384. In one specific embodiment, the antibody comprisesthe same VL and VH CDRs as G9.1-9.

In one specific embodiment, the anti-Galectin-9 antibody or bindingportion thereof comprises heavy and light chain variable regions,wherein: the light chain variable region CDR1, CDR2, and CDR3 compriseSEQ ID NO: 328, 329, and 339, respectively, and the heavy chain variableregion CDR1, CDR2, and CDR3 comprise SEQ ID NO: 429, 444, and 385. Insome embodiments, the light and heavy chain variable region CDR1, CDR2,and CDR3 regions consist of SEQ ID NO: 328, 329, and 339, and SEQ ID NO:429, 444, and 385. In one specific embodiment, the antibody comprisesthe same VL and VH CDRs as G9.1-10.

In one specific embodiment, the anti-Galectin-9 antibody or bindingportion thereof comprises heavy and light chain variable regions,wherein: the light chain variable region CDR1, CDR2, and CDR3 compriseSEQ ID NO: 328, 329, and 340, respectively, and the heavy chain variableregion CDR1, CDR2, and CDR3 comprise SEQ ID NO: 428, 445, and 386. Insome embodiments, the light and heavy chain variable region CDR1, CDR2,and CDR3 regions consist of SEQ ID NO: 328, 329, and 340, and SEQ ID NO:428, 445, and 386. In one specific embodiment, the antibody comprisesthe same VL and VH CDRs as G9.1-11.

In one specific embodiment, the anti-Galectin-9 antibody or bindingportion thereof comprises heavy and light chain variable regions,wherein: the light chain variable region CDR1, CDR2, and CDR3 compriseSEQ ID NO: 328, 329, and 337, respectively, and the heavy chain variableregion CDR1, CDR2, and CDR3 comprise SEQ ID NO: 361, 365, and 374. Insome embodiments, the light and heavy chain variable region CDR1, CDR2,and CDR3 regions consist of SEQ ID NO: 328, 329, and 337, and SEQ ID NO:361, 365, and 374. In one specific embodiment, the antibody comprisesthe same VL and VH CDRs as G9.1-8m1.

In one specific embodiment, the anti-Galectin-9 antibody or bindingportion thereof comprises heavy and light chain variable regions,wherein: the light chain variable region CDR1, CDR2, and CDR3 compriseSEQ ID NO: 328, 329, and 337, respectively, and the heavy chain variableregion CDR1, CDR2, and CDR3 comprise SEQ ID NO: 361, 366, and 374. Insome embodiments, the light and heavy chain variable region CDR1, CDR2,and CDR3 regions consist of SEQ ID NO: 328, 329, and 337, and SEQ ID NO:361, 366, and 374. In one specific embodiment, the antibody comprisesthe same VL and VH CDRs as G9.1-8m2.

In one specific embodiment, the anti-Galectin-9 antibody or bindingportion thereof comprises heavy and light chain variable regions,wherein: the light chain variable region CDR1, CDR2, and CDR3 compriseSEQ ID NO: 328, 329, and 337, respectively, and the heavy chain variableregion CDR1, CDR2, and CDR3 comprise SEQ ID NO: 361, 364, and 375. Insome embodiments, the light and heavy chain variable region CDR1, CDR2,and CDR3 regions consist of SEQ ID NO: 328, 329, and 337, and SEQ ID NO:361, 364, and 375. In one specific embodiment, the antibody comprisesthe same VL and VH CDRs as G9.1-8m3.

In one specific embodiment, the anti-Galectin-9 antibody or bindingportion thereof comprises heavy and light chain variable regions,wherein: the light chain variable region CDR1, CDR2, and CDR3 compriseSEQ ID NO: 328, 329, and 337, respectively, and the heavy chain variableregion CDR1, CDR2, and CDR3 comprise SEQ ID NO: 361, 364, and 376. Insome embodiments, the light and heavy chain variable region CDR1, CDR2,and CDR3 regions consist of SEQ ID NO: 328, 329, and 337, and SEQ ID NO:361, 364, and 376. In one specific embodiment, the antibody comprisesthe same VL and VH CDRs as G9.1-8m4.

In one specific embodiment, the anti-Galectin-9 antibody or bindingportion thereof comprises heavy and light chain variable regions,wherein: the light chain variable region CDR1, CDR2, and CDR3 compriseSEQ ID NO: 328, 329, and 337, respectively, and the heavy chain variableregion CDR1, CDR2, and CDR3 comprise SEQ ID NO: 361, 364, and 377. Insome embodiments, the light and heavy chain variable region CDR1, CDR2,and CDR3 regions consist of SEQ ID NO: 328, 329, and 337, and SEQ ID NO:361, 364, and 377. In one specific embodiment, the antibody comprisesthe same VL and VH CDRs as G9.1-8m5.

In one specific embodiment, the anti-Galectin-9 antibody or bindingportion thereof comprises heavy and light chain variable regions,wherein: the light chain variable region CDR1, CDR2, and CDR3 compriseSEQ ID NO: 328, 329, and 337, respectively, and the heavy chain variableregion CDR1, CDR2, and CDR3 comprise SEQ ID NO: 361, 364, and 378. Insome embodiments, the light and heavy chain variable region CDR1, CDR2,and CDR3 regions consist of SEQ ID NO: 328, 329, and 337, and SEQ ID NO:361, 364, and 378. In one specific embodiment, the antibody comprisesthe same VL and VH CDRs as G9.1-8m6.

In one specific embodiment, the anti-Galectin-9 antibody or bindingportion thereof comprises heavy and light chain variable regions,wherein: the light chain variable region CDR1, CDR2, and CDR3 compriseSEQ ID NO: 328, 329, and 337, respectively, and the heavy chain variableregion CDR1, CDR2, and CDR3 comprise SEQ ID NO: 361, 364, and 379. Insome embodiments, the light and heavy chain variable region CDR1, CDR2,and CDR3 regions consist of SEQ ID NO: 328, 329, and 337, and SEQ ID NO:361, 364, and 379. In one specific embodiment, the antibody comprisesthe same VL and VH CDRs as G9.1-8m7.

In one specific embodiment, the anti-Galectin-9 antibody or bindingportion thereof comprises heavy and light chain variable regions,wherein: the light chain variable region CDR1, CDR2, and CDR3 compriseSEQ ID NO: 328, 329, and 337, respectively, and the heavy chain variableregion CDR1, CDR2, and CDR3 comprise SEQ ID NO: 361, 364, and 380. Insome embodiments, the light and heavy chain variable region CDR1, CDR2,and CDR3 regions consist of SEQ ID NO: 328, 329, and 337, and SEQ ID NO:361, 364, and 380. In one specific embodiment, the antibody comprisesthe same VL and VH CDRs as G9.1-8m8.

In one specific embodiment, the anti-Galectin-9 antibody or bindingportion thereof comprises heavy and light chain variable regions,wherein: the light chain variable region CDR1, CDR2, and CDR3 compriseSEQ ID NO: 328, 329, and 337, respectively, and the heavy chain variableregion CDR1, CDR2, and CDR3 comprise SEQ ID NO: 361, 364, and 383. Insome embodiments, the light and heavy chain variable region CDR1, CDR2,and CDR3 regions consist of SEQ ID NO: 328, 329, and 337, and SEQ ID NO:361, 364, and 383. In one specific embodiment, the antibody comprisesthe same VL and VH CDRs as G9.1-8m9.

In one specific embodiment, the anti-Galectin-9 antibody or bindingportion thereof comprises heavy and light chain variable regions,wherein: the light chain variable region CDR1, CDR2, and CDR3 compriseSEQ ID NO: 328, 329, and 337, respectively, and the heavy chain variableregion CDR1, CDR2, and CDR3 comprise SEQ ID NO: 361, 364, and 381. Insome embodiments, the light and heavy chain variable region CDR1, CDR2,and CDR3 regions consist of SEQ ID NO: 328, 329, and 337, and SEQ ID NO:361, 364, and 381. In one specific embodiment, the antibody comprisesthe same VL and VH CDRs as G9.1-8m10.

In one specific embodiment, the anti-Galectin-9 antibody or bindingportion thereof comprises heavy and light chain variable regions,wherein: the light chain variable region CDR1, CDR2, and CDR3 compriseSEQ ID NO: 328, 329, and 337, respectively, and the heavy chain variableregion CDR1, CDR2, and CDR3 comprise SEQ ID NO: 361, 364, and 382. Insome embodiments, the light and heavy chain variable region CDR1, CDR2,and CDR3 regions consist of SEQ ID NO: 328, 329, and 337, and SEQ ID NO:361, 364, and 382. In one specific embodiment, the antibody comprisesthe same VL and VH CDRs as G9.1-8m11.

In one specific embodiment, the anti-Galectin-9 antibody or bindingportion thereof comprises heavy and light chain variable regions,wherein: the light chain variable region CDR1, CDR2, and CDR3 compriseSEQ ID NO: 328, 329, and 337, respectively, and the heavy chain variableregion CDR1, CDR2, and CDR3 comprise SEQ ID NO: 361, 366, and 380. Insome embodiments, the light and heavy chain variable region CDR1, CDR2,and CDR3 regions consist of SEQ ID NO: 328, 329, and 337, and SEQ ID NO:361, 366, and 380. In one specific embodiment, the antibody comprisesthe same VL and VH CDRs as G9.1-8m12.

In one specific embodiment, the anti-Galectin-9 antibody or bindingportion thereof comprises heavy and light chain variable regions,wherein: the light chain variable region CDR1, CDR2, and CDR3 compriseSEQ ID NO: 328, 329, and 337, respectively, and the heavy chain variableregion CDR1, CDR2, and CDR3 comprise SEQ ID NO: 361, 366, and 383. Insome embodiments, the light and heavy chain variable region CDR1, CDR2,and CDR3 regions consist of SEQ ID NO: 328, 329, and 337, and SEQ ID NO:361, 366, and 383.

In one specific embodiment, the antibody comprises the same VL and VHCDRs as G9.1-8m13. In one specific embodiment, the anti-Galectin-9antibody or binding portion thereof comprises heavy and light chainvariable regions, wherein: the light chain variable region CDR1, CDR2,and CDR3 comprise SEQ ID NO: 328, 329, and 337, respectively, and theheavy chain variable region CDR1, CDR2, and CDR3 comprise SEQ ID NO:361, 366, and 382, respectively. In some embodiments, the light andheavy chain variable region CDR1, CDR2, and CDR3 regions consist of SEQID NO: 328, 329, and 337, and SEQ ID NO: 361, 366, and 382. In onespecific embodiment, the antibody comprises the same VL and VH CDRs asG9.1-8m14.

Sequence Identity

In some embodiments, the anti-Galectin-9 antibody (e.g., specific toCRD1 and/or CRD2) may comprise light chain CDRs that have at least 80%(e.g., 85%, 90%, 95%, 96%, 97%, 98%, 99% and any increment therein)sequence identity, individually or collectively, as compared with thecorresponding VL CDRs of an exemplary antibody described herein.Alternatively or in addition, the anti-Galectin-9 antibody (e.g.,specific to CRD1 or CRD2) may comprise heavy chain CDRs that have atleast 80% (e.g., 85%, 90%, 95%, 96%, 97%, 98%, 99% and any incrementtherein) sequence identity, individually or collectively, as comparedwith the V_(H) CDRs as an exemplary antibody described herein.

In some embodiments, the anti-Galectin-9 antibody (e.g., specific toCRD1 and/or CRD2) may comprise light chain CDRs that have at least 80%(e.g., 85%, 90%, 95%, 96%, 97%, 98%, 99% and any increment therein)sequence identity, individually or collectively, as compared with thecorresponding VL CDRs of an antibody or antigen binding portion thereofselected from G9.1-1, G9.1-2, G9.1-3, G9.1-4, G9.1-5, G9.1-6, G9.1-7,G9.1-8, G9.1-9, G9.1-10, G9.1-11, G9.1-8m1, G9.1-8m2, G9.1-8m3,G9.1-8m4, G9.1-8m5, G9.1-8m6, G9.1-8m7, G9.1-8m8, G9.1-8m9, G9.1-8m10,G9.1-8m11, G9.1-8m12, G9.1-8m13, and G9.1-8m14. In some embodiments, theanti-Galectin-9 antibody (e.g., specific to CRD1 and/or CRD2) maycomprise heavy chain CDRs that have at least 80% (e.g., 85%, 90%, 95%,96%, 97%, 98%, 99% and any increment therein) sequence identity,individually or collectively, as compared with the corresponding VH CDRsof an antibody or antigen binding portion thereof selected from G9.1-1,G9.1-2, G9.1-3, G9.1-4, G9.1-5, G9.1-6, G9.1-7, G9.1-8, G9.1-9, G9.1-10,G9.1-11, G9.1-8m1, G9.1-8m2, G9.1-8m3, G9.1-8m4, G9.1-8m5, G9.1-8m6,G9.1-8m7, G9.1-8m8, G9.1-8m9, G9.1-8m10, G9.1-8m11, G9.1-8m12,G9.1-8m13, and G9.1-8m14.

In some embodiments, the anti-Galectin-9 antibody (e.g., specific toCRD1 and/or CRD2) may comprise light chain CDRs and heavy chain CDRsthat have at least 80% (e.g., 85%, 90%, 95%, 96%, 97%, 98%, 99% and anyincrement therein) sequence identity, individually or collectively, ascompared with the corresponding VL CDRs and VH CDRs of an antibody orantigen binding portion thereof selected from G9.1-1, G9.1-2, G9.1-3,G9.1-4, G9.1-5, G9.1-6, G9.1-7, G9.1-8, G9.1-9, G9.1-10, G9.1-11,G9.1-8m1, G9.1-8m2, G9.1-8m3, G9.1-8m4, G9.1-8m5, G9.1-8m6, G9.1-8m7,G9.1-8m8, G9.1-8m9, G9.1-8m10, G9.1-8m11, G9.1-8m12, G9.1-8m13, andG9.1-8m14.

In some embodiments, the anti-Galectin-9 antibody (e.g., specific toCRD1 and/or CRD2) may comprise a VL CDR1 amino acid sequence that has atleast 80% (e.g., 85%, 90%, 95%, 96%, 97%, 98%, 99% and any incrementtherein) sequence identity to VL CDR1 amino acid sequence set forth inSEQ ID NO: 374. In some embodiments, the anti-Galectin-9 antibody (e.g.,specific to CRD1 and/or CRD2) may comprise a VL CDR2 amino acid sequencethat has at least 80% (e.g., 85%, 90%, 95%, 96%, 97%, 98%, 99% and anyincrement therein) sequence identity to the VL CDR2 amino acid sequenceset forth in SEQ ID NO: 329. In some embodiments, the anti-Galectin-9antibody (e.g., specific to CRD1 and/or CRD2) may comprise a VL CDR3amino acid sequence that has at least 80% (e.g., 85%, 90%, 95%, 96%,97%, 98%, 99% and any increment therein) sequence identity to a VL CDR3amino acid sequence selected from SEQ ID NO: 330-340.

In some embodiments, the anti-Galectin-9 antibody (e.g., specific toCRD1 and/or CRD2) may comprise a VH CDR1 amino acid sequence that has atleast 80% (e.g., 85%, 90%, 95%, 96%, 97%, 98%, 99% and any incrementtherein) sequence identity to a VH CDR1 amino acid sequence set forth inSEQ ID NO: 361, 427, 428, 431, 435, 436, 437. In some embodiments, theanti-Galectin-9 antibody (e.g., specific to CRD1 and/or CRD2) maycomprise a VH CDR2 amino acid sequence that has at least 80% (e.g., 85%,90%, 95%, 96%, 97%, 98%, 99% and any increment therein) sequenceidentity to a VH CDR2 amino acid sequence selected from SEQ ID NO:362-366 and 438-445. In some embodiments, the anti-Galectin-9 antibody(e.g., specific to CRD1 and/or CRD2) may comprise a VH CDR3 amino acidsequence that has at least 80% (e.g., 85%, 90%, 95%, 96%, 97%, 98%, 99%and any increment therein) sequence identity to a VH CDR3 amino acidsequence selected from SEQ ID NO: 367-386.

Accordingly, in some embodiments, anti-Galectin-9 antibodies or antigenbinding portions thereof comprise (a) VL CDR1 amino acid sequence thathas at least 80% (e.g., 85%, 90%, 95%, 96%, 97%, 98%, 99% and anyincrement therein) sequence identity to the VL CDR1 amino acid sequenceset forth in SEQ ID NO: 328; (b) VL CDR2 amino acid sequence that has atleast 80% (e.g., 85%, 90%, 95%, 96%, 97%, 98%, 99% and any incrementtherein) sequence identity to the VL CDR2 amino acid sequence set forthin SEQ ID NO: 329; (c) VL CDR3 amino acid sequence that has at least 80%(e.g., 85%, 90%, 95%, 96%, 97%, 98%, 99% and any increment therein)sequence identity to a VL CDR3 amino acid sequence selected from SEQ IDNO: 330-340; (d) VH CDR1 amino acid sequence that has at least 80%(e.g., 85%, 90%, 95%, 96%, 97%, 98%, 99% and any increment therein)sequence identity to the VH CDR1 amino acid sequence set forth in SEQ IDNO: 361, 427, 428, 431, 435, 436, 437; (e) VH CDR2 amino acid sequencethat has at least 80% (e.g., 85%, 90%, 95%, 96%, 97%, 98%, 99% and anyincrement therein) sequence identity to a VH CDR2 amino acid sequenceselected from SEQ ID NO: 362-366 and 438-445; (f) VH CDR3 amino acidsequence that has at least 80% (e.g., 85%, 90%, 95%, 96%, 97%, 98%, 99%and any increment therein) sequence identity to a VH CDR3 amino acidsequence selected from SEQ ID NO: 367-386.

In some embodiments, the anti-Galectin-9 antibody or binding portionthereof comprises heavy and light chain variable regions, wherein thelight chain variable region CDR1, CDR2, and CDR3 amino acid sequenceshave at least 80% (e.g., 85%, 90%, 95%, 96%, 97%, 98%, 99% and anyincrement therein) sequence identity to the light chain variable regionCDR1, CDR2, and CDR3 amino acid sequences set forth in SEQ ID NO: 328,329, and 337, respectively. In some embodiments, the antibody VL CDR1,CDR2, and CDR3 amino acid sequences have at least 80% (e.g., 85%, 90%,95%, 96%, 97%, 98%, 99% and any increment therein) sequence identity tothe VL CDR1, CDR2, and CDR3 amino acid sequences of G9.1-8m13. In someembodiments, the anti-Galectin-9 antibody or binding portion thereofcomprises heavy and light chain variable regions, wherein the heavychain variable region CDR1, CDR2, and CDR3 amino acid sequences have atleast 80% (e.g., 85%, 90%, 95%, 96%, 97%, 98%, 99% and any incrementtherein) sequence identity to the heavy chain variable region CDR1,CDR2, and CDR3 amino acid sequences set forth in SEQ ID NO: 361, 366,and 383. In some embodiments, the antibody VH CDR1, CDR2, and CDR3 aminoacid sequences have at least 80% (e.g., 85%, 90%, 95%, 96%, 97%, 98%,99% and any increment therein) sequence identity to the VH CDR1, CDR2,and CDR3 amino acid sequences of G9.1-8m13. In one specific embodiment,the anti-Galectin-9 antibody or binding portion thereof comprises heavyand light chain variable regions, wherein: the light chain variableregion CDR1, CDR2, and CDR3 amino acid sequences have at least 80%(e.g., 85%, 90%, 95%, 96%, 97%, 98%, 99% and any increment therein)sequence identity to the light chain variable region CDR1, CDR2, andCDR3 amino acid sequences set forth in SEQ ID NO: 328, 329, and 337,respectively, and the heavy chain variable region CDR1, CDR2, and CDR3amino acid sequences have at least 80% (e.g., 85%, 90%, 95%, 96%, 97%,98%, 99% and any increment therein) sequence identity to the heavy chainvariable region CDR1, CDR2, and CDR3 amino acid sequences set forth inSEQ ID NO: 361, 366, and 383. In one specific embodiment, the antibodyVL CDR1, CDR2, and CDR3 and VH CDR1, CDR2, and CDR3 amino acid sequenceshave at least 80% (e.g., 85%, 90%, 95%, 96%, 97%, 98%, 99% and anyincrement therein) sequence identity to the VL CDR1, CDR2, and CDR3 andVH CDR1, CDR2, and CDR3 amino acid sequences as G9.1-8m13.

9.2 Antibody Clones and Related CDRs

Clone 9.2-Derived Light Chain Variable Region

In some embodiments, the anti-Galectin-9 antibody or binding portionthereof comprises heavy and light chain variable regions, wherein thelight chain variable region CDR1, CDR2, and CDR3 regions comprise SEQ IDNO: 328, 329, and 341, respectively. In some embodiments, the lightchain variable region CDR1, CDR2, and CDR3 regions consist of SEQ ID NO:328, 329, and 341, respectively. In some embodiments, the antibodycomprises the same VL CDRs as G9.2-1.

In some embodiments, the anti-Galectin-9 antibody or binding portionthereof comprises heavy and light chain variable regions, wherein thelight chain variable region CDR1, CDR2, and CDR3 regions comprise SEQ IDNO: 328, 329, and 333, respectively. In some embodiments, the lightchain variable region CDR1, CDR2, and CDR3 regions consist of SEQ ID NO:328, 329, and 333, respectively. In some embodiments, the antibodycomprises the same VL CDRs as G9.2-2.

In some embodiments, the anti-Galectin-9 antibody or binding portionthereof comprises heavy and light chain variable regions, wherein thelight chain variable region CDR1, CDR2, and CDR3 regions comprise SEQ IDNO: 328, 329, and 333, respectively. In some embodiments, the lightchain variable region CDR1, CDR2, and CDR3 regions consist of SEQ ID NO:328, 329, and 333, respectively. In some embodiments, the antibodycomprises the same VL CDRs as G9.2-3.

In some embodiments, the anti-Galectin-9 antibody or binding portionthereof comprises heavy and light chain variable regions, wherein thelight chain variable region CDR1, CDR2, and CDR3 regions comprise SEQ IDNO: 328, 329, and 342, respectively. In some embodiments, the lightchain variable region CDR1, CDR2, and CDR3 regions consist of SEQ ID NO:328, 329, and 342, respectively. In some embodiments, the antibodycomprises the same VL CDRs as G9.2-4.

In some embodiments, the anti-Galectin-9 antibody or binding portionthereof comprises heavy and light chain variable regions, wherein thelight chain variable region CDR1, CDR2, and CDR3 regions comprise SEQ IDNO: 328, 329, and 343, respectively. In some embodiments, the lightchain variable region CDR1, CDR2, and CDR3 regions consist of SEQ ID NO:328, 329, and 343, respectively. In some embodiments, the antibodycomprises the same VL CDRs as G9.2-5.

In some embodiments, the anti-Galectin-9 antibody or binding portionthereof comprises heavy and light chain variable regions, wherein thelight chain variable region CDR1, CDR2, and CDR3 regions comprise SEQ IDNO: 328, 329, and 344, respectively. In some embodiments, the lightchain variable region CDR1, CDR2, and CDR3 regions consist of SEQ ID NO:328, 329, and 344, respectively. In some embodiments, the antibodycomprises the same VL CDRs as G9.2-6.

In some embodiments, the anti-Galectin-9 antibody or binding portionthereof comprises heavy and light chain variable regions, wherein thelight chain variable region CDR1, CDR2, and CDR3 regions comprise SEQ IDNO: 328, 329, and 345, respectively. In some embodiments, the lightchain variable region CDR1, CDR2, and CDR3 regions consist of SEQ ID NO:328, 329, and 345, respectively. In some embodiments, the antibodycomprises the same VL CDRs as G9.2-7. In some embodiments, theanti-Galectin-9 antibody or binding portion thereof comprises heavy andlight chain variable regions, wherein the light chain variable regionCDR1, CDR2, and CDR3 regions comprise SEQ ID NO: 328, 329, and 346,respectively. In some embodiments, the light chain variable region CDR1,CDR2, and CDR3 regions consist of SEQ ID NO: 328, 329, and 346. In someembodiments, the antibody comprises the same VL CDRs as G9.2-8.

In some embodiments, the anti-Galectin-9 antibody or binding portionthereof comprises heavy and light chain variable regions, wherein thelight chain variable region CDR1, CDR2, and CDR3 regions comprise SEQ IDNO: 328, 329, and 347, respectively. In some embodiments, the lightchain variable region CDR1, CDR2, and CDR3 regions consist of SEQ ID NO:328, 329, and 347, respectively. In some embodiments, the antibodycomprises the same VL CDRs as G9.2-9.

In some embodiments, the anti-Galectin-9 antibody or binding portionthereof comprises heavy and light chain variable regions, wherein thelight chain variable region CDR1, CDR2, and CDR3 regions comprise SEQ IDNO: 328, 329, and 348, respectively. In some embodiments, the lightchain variable region CDR1, CDR2, and CDR3 regions consist of SEQ ID NO:328, 329, and 348, respectively. In some embodiments, the antibodycomprises the same VL CDRs as G9.2-10.

In some embodiments, the anti-Galectin-9 antibody or binding portionthereof comprises heavy and light chain variable regions, wherein thelight chain variable region CDR1, CDR2, and CDR3 regions comprise SEQ IDNO: 328, 329, and 349, respectively. In some embodiments, the lightchain variable region CDR1, CDR2, and CDR3 regions consist of SEQ ID NO:328, 329, and 349, respectively. In some embodiments, the antibodycomprises the same VL CDRs as G9.2-11.

In some embodiments, the anti-Galectin-9 antibody or binding portionthereof comprises heavy and light chain variable regions, wherein thelight chain variable region CDR1, CDR2, and CDR3 regions comprise SEQ IDNO: 328, 329, and 350, respectively. In some embodiments, the lightchain variable region CDR1, CDR2, and CDR3 regions consist of SEQ ID NO:328, 329, and 350, respectively. In some embodiments, the antibodycomprises the same VL CDRs as G9.2-12.

In some embodiments, the anti-Galectin-9 antibody or binding portionthereof comprises heavy and light chain variable regions, wherein thelight chain variable region CDR1, CDR2, and CDR3 regions comprise SEQ IDNO: 328, 329, and 341, respectively. In some embodiments, the lightchain variable region CDR1, CDR2, and CDR3 regions consist of SEQ ID NO:328, 329, and 341. In some embodiments, the antibody comprises the sameVL CDRs as G9.2-13.

In some embodiments, the anti-Galectin-9 antibody or binding portionthereof comprises heavy and light chain variable regions, wherein thelight chain variable region CDR1, CDR2, and CDR3 regions comprise SEQ IDNO: 328, 329, and 333, respectively. In some embodiments, the lightchain variable region CDR1, CDR2, and CDR3 regions consist of SEQ ID NO:328, 329, and 333, respectively. In some embodiments, the antibodycomprises the same VL CDRs as G9.2-14.

In some embodiments, the anti-Galectin-9 antibody or binding portionthereof comprises heavy and light chain variable regions, wherein thelight chain variable region CDR1, CDR2, and CDR3 regions comprise SEQ IDNO: 328, 329, and 343, respectively. In some embodiments, the lightchain variable region CDR1, CDR2, and CDR3 regions consist of SEQ ID NO:328, 329, and 343, respectively. In some embodiments, the antibodycomprises the same VL CDRs as G9.2-15.

In some embodiments, the anti-Galectin-9 antibody or binding portionthereof comprises heavy and light chain variable regions, wherein thelight chain variable region CDR1, CDR2, and CDR3 regions comprise SEQ IDNO: 328, 329, and 333, respectively. In some embodiments, the lightchain variable region CDR1, CDR2, and CDR3 regions consist of SEQ ID NO:328, 329, and 333. In some embodiments, the antibody comprises the sameVL CDRs as G9.2-16.

In some embodiments, the anti-Galectin-9 antibody or binding portionthereof comprises heavy and light chain variable regions, wherein thelight chain variable region CDR1 comprises SEQ ID NO: 328. In someembodiments, the anti-Galectin-9 antibody or binding portion thereofcomprises heavy and light chain variable regions, wherein the lightchain variable region CDR2 comprises SEQ ID NO: 329. In someembodiments, the anti-Galectin-9 antibody or binding portion thereofcomprises heavy and light chain variable regions, wherein the lightchain variable region CDR3 comprises SEQ ID NO: 352. In someembodiments, the anti-Galectin-9 antibody or binding portion thereofcomprises heavy and light chain variable regions, wherein the lightchain variable region CDR1 consists of SEQ ID NO: 328. In someembodiments, the anti-Galectin-9 antibody or binding portion thereofcomprises heavy and light chain variable regions, wherein the lightchain variable region CDR2 consists of SEQ ID NO: 329. In someembodiments, the anti-Galectin-9 antibody or binding portion thereofconsists of heavy and light chain variable regions, wherein the lightchain variable region CDR3 comprises SEQ ID NO: 352. In someembodiments, the anti-Galectin-9 antibody or binding portion thereofcomprises heavy and light chain variable regions, wherein the lightchain variable region CDR1, CDR2, and CDR3 regions comprise SEQ ID NO:328, 329, and 352, respectively. In some embodiments, the light chainvariable region CDR1, CDR2, and CDR3 regions consist of SEQ ID NO: 328,329, and 352, respectively. In some embodiments, the antibody comprisesthe same VL CDRs as G9.2-17.

In some embodiments, the anti-Galectin-9 antibody or binding portionthereof comprises heavy and light chain variable regions, wherein thelight chain variable region CDR1, CDR2, and CDR3 regions comprise SEQ IDNO: 328, 329, and 352, respectively. In some embodiments, the lightchain variable region CDR1, CDR2, and CDR3 regions consist of SEQ ID NO:328, 329, and 352, respectively. In some embodiments, the antibodycomprises the same VL CDRs as G9.2-17mut6.

In some embodiments, the anti-Galectin-9 antibody or binding portionthereof comprises heavy and light chain variable regions, wherein thelight chain variable region CDR1, CDR2, and CDR3 regions comprise SEQ IDNO: 328, 329, and 333, respectively. In some embodiments, the lightchain variable region CDR1, CDR2, and CDR3 regions consist of SEQ ID NO:328, 329, and 333, respectively. In some embodiments, the antibodycomprises the same VL CDRs as G9.2-18.

In some embodiments, the anti-Galectin-9 antibody or binding portionthereof comprises heavy and light chain variable regions, wherein thelight chain variable region CDR1, CDR2, and CDR3 regions comprise SEQ IDNO: 328, 329, and 354, respectively. In some embodiments, the lightchain variable region CDR1, CDR2, and CDR3 regions consist of SEQ ID NO:328, 329, and 354. In some embodiments, the antibody comprises the sameVL CDRs as G9.2-19.

In some embodiments, the anti-Galectin-9 antibody or binding portionthereof comprises heavy and light chain variable regions, wherein thelight chain variable region CDR1, CDR2, and CDR3 regions comprise SEQ IDNO: 328, 329, and 352, respectively. In some embodiments, the lightchain variable region CDR1, CDR2, and CDR3 regions consist of SEQ ID NO:328, 329, and 352, respectively. In some embodiments, the antibodycomprises the same VL CDRs as G9.2-20.

In some embodiments, the anti-Galectin-9 antibody or binding portionthereof comprises heavy and light chain variable regions, wherein thelight chain variable region CDR1, CDR2, and CDR3 regions comprise SEQ IDNO: 328, 329, and 355, respectively. In some embodiments, the lightchain variable region CDR1, CDR2, and CDR3 regions consist of SEQ ID NO:328, 329, and 355, respectively. In some embodiments, the antibodycomprises the same VL CDRs as G9.2-21.

In some embodiments, the anti-Galectin-9 antibody or binding portionthereof comprises heavy and light chain variable regions, wherein thelight chain variable region CDR1, CDR2, and CDR3 regions comprise SEQ IDNO: 328, 329, and 356, respectively. In some embodiments, the lightchain variable region CDR1, CDR2, and CDR3 regions consist of SEQ ID NO:328, 329, and 356, respectively. In some embodiments, the antibodycomprises the same VL CDRs as G9.2-22.

In some embodiments, the anti-Galectin-9 antibody or binding portionthereof comprises heavy and light chain variable regions, wherein thelight chain variable region CDR1, CDR2, and CDR3 regions comprise SEQ IDNO: 328, 329, and 357, respectively. In some embodiments, the lightchain variable region CDR1, CDR2, and CDR3 regions consist of SEQ ID NO:328, 329, and 357, respectively. In some embodiments, the antibodycomprises the same VL CDRs as G9.2-23.

In some embodiments, the anti-Galectin-9 antibody or binding portionthereof comprises heavy and light chain variable regions, wherein thelight chain variable region CDR1, CDR2, and CDR3 regions comprise SEQ IDNO: 328, 329, and 358, respectively. In some embodiments, the lightchain variable region CDR1, CDR2, and CDR3 regions consist of SEQ ID NO:328, 329, and 358, respectively. In some embodiments, the antibodycomprises the same VL CDRs as G9.2-24.

In some embodiments, the anti-Galectin-9 antibody or binding portionthereof comprises heavy and light chain variable regions, wherein thelight chain variable region CDR1, CDR2, and CDR3 regions comprise SEQ IDNO: 328, 329, and 359, respectively. In some embodiments, the lightchain variable region CDR1, CDR2, and CDR3 regions consist of SEQ ID NO:328, 329, and 359, respectively. In some embodiments, the antibodycomprises the same VL CDRs as G9.2-25.

In some embodiments, the anti-Galectin-9 antibody or binding portionthereof comprises heavy and light chain variable regions, wherein thelight chain variable region CDR1, CDR2, and CDR3 regions comprise SEQ IDNO: 328, 329, and 360, respectively. In some embodiments, the lightchain variable region CDR1, CDR2, and CDR3 regions consist of SEQ ID NO:328, 329, and 360, respectively. In some embodiments, the antibodycomprises the same VL CDRs as G9.2-26.

In some embodiments, the anti-Galectin-9 antibody or binding portionthereof comprises heavy and light chain variable regions, wherein thelight chain variable region CDR1, CDR2, and CDR3 regions comprise SEQ IDNO: 328, 329, and 352, respectively. In some embodiments, the lightchain variable region CDR1, CDR2, and CDR3 regions consist of SEQ ID NO:328, 329, and 352, respectively. In some embodiments, the antibodycomprises the same VL CDRs as G9.2-low affinity binder.

Clone 9.2-Derived Heavy Chain Variable Region

In some embodiments, the anti-Galectin-9 antibody or binding portionthereof comprises heavy and light chain variable regions, wherein theheavy chain variable region CDR1, CDR2, and CDR3 regions comprise SEQ IDNO: 424, 446, and 390, respectively. In some embodiments, the heavychain variable region CDR1, CDR2, and CDR3 regions consist of SEQ ID NO:424, 446, and 390, respectively. In some embodiments, the antibodycomprises the same VH CDRs as G9.2-1.

In some embodiments, the anti-Galectin-9 antibody or binding portionthereof comprises heavy and light chain variable regions, wherein theheavy chain variable region CDR1, CDR2, and CDR3 regions comprise SEQ IDNO: 431, 447, and 391, respectively. In some embodiments, the heavychain variable region CDR1, CDR2, and CDR3 regions consist of SEQ ID NO:431, 447, and 391, respectively. In some embodiments, the antibodycomprises the same VH CDRs as G9.2-2.

In some embodiments, the anti-Galectin-9 antibody or binding portionthereof comprises heavy and light chain variable regions, wherein theheavy chain variable region CDR1, CDR2, and CDR3 regions comprise SEQ IDNO: 431, 448, and 392, respectively. In some embodiments, the heavychain variable region CDR1, CDR2, and CDR3 regions consist of SEQ ID NO:431, 448, and 392. In some embodiments, the antibody comprises the sameVH CDRs as G9.2-3.

In some embodiments, the anti-Galectin-9 antibody or binding portionthereof comprises heavy and light chain variable regions, wherein theheavy chain variable region CDR1, CDR2, and CDR3 regions comprise SEQ IDNO: 431, 449, and 393, respectively. In some embodiments, the heavychain variable region CDR1, CDR2, and CDR3 regions consist of SEQ ID NO:431, 449, and 393, respectively. In some embodiments, the antibodycomprises the same VH CDRs as G9.2-4.

In some embodiments, the anti-Galectin-9 antibody or binding portionthereof comprises heavy and light chain variable regions, wherein theheavy chain variable region CDR1, CDR2, and CDR3 regions comprise SEQ IDNO: 431, 450, and 394, respectively. In some embodiments, the heavychain variable region CDR1, CDR2, and CDR3 regions consist of SEQ ID NO:431, 450, and 394, respectively. In some embodiments, the antibodycomprises the same VH CDRs as G9.2-5.

In some embodiments, the anti-Galectin-9 antibody or binding portionthereof comprises heavy and light chain variable regions, wherein theheavy chain variable region CDR1, CDR2, and CDR3 regions comprise SEQ IDNO: 431, 451, and 395, respectively. In some embodiments, the heavychain variable region CDR1, CDR2, and CDR3 regions consist of SEQ ID NO:431, 452, and 395. In some embodiments, the antibody comprises the sameVH CDRs as G9.2-6.

In some embodiments, the anti-Galectin-9 antibody or binding portionthereof comprises heavy and light chain variable regions, wherein theheavy chain variable region CDR1, CDR2, and CDR3 regions comprise SEQ IDNO: 425, 453, and 396, respectively. In some embodiments, the heavychain variable region CDR1, CDR2, and CDR3 regions consist of SEQ ID NO:425, 453, and 396, respectively. In some embodiments, the antibodycomprises the same VH CDRs as G9.2-7.

In some embodiments, the anti-Galectin-9 antibody or binding portionthereof comprises heavy and light chain variable regions, wherein theheavy chain variable region CDR1, CDR2, and CDR3 regions comprise SEQ IDNO: 425, 453, and 397, respectively. In some embodiments, the heavychain variable region CDR1, CDR2, and CDR3 regions consist of SEQ ID NO:425, 453, and 397, respectively. In some embodiments, the antibodycomprises the same VH CDRs as G9.2-8.

In some embodiments, the anti-Galectin-9 antibody or binding portionthereof comprises heavy and light chain variable regions, wherein theheavy chain variable region CDR1, CDR2, and CDR3 regions comprise SEQ IDNO: 426, 454, and 398, respectively. In some embodiments, the heavychain variable region CDR1, CDR2, and CDR3 regions consist of SEQ ID NO:426, 454, and 398, respectively. In some embodiments, the antibodycomprises the same VH CDRs as G9.2-9.

In some embodiments, the anti-Galectin-9 antibody or binding portionthereof comprises heavy and light chain variable regions, wherein theheavy chain variable region CDR1, CDR2, and CDR3 regions comprise SEQ IDNO: 426, 387, and 399, respectively. In some embodiments, the heavychain variable region CDR1, CDR2, and CDR3 regions consist of SEQ ID NO:426, 387, and 399, respectively. In some embodiments, the antibodycomprises the same VH CDRs as G9.2-10.

In some embodiments, the anti-Galectin-9 antibody or binding portionthereof comprises heavy and light chain variable regions, wherein theheavy chain variable region CDR1, CDR2, and CDR3 regions comprise SEQ IDNO: 432, 455, and 400, respectively. In some embodiments, the heavychain variable region CDR1, CDR2, and CDR3 regions consist of SEQ ID NO:432, 455, and 400, respectively. In some embodiments, the antibodycomprises the same VH CDRs as G9.2-11.

In some embodiments, the anti-Galectin-9 antibody or binding portionthereof comprises heavy and light chain variable regions, wherein theheavy chain variable region CDR1, CDR2, and CDR3 regions comprise SEQ IDNO: 433, 456, and 401, respectively. In some embodiments, the heavychain variable region CDR1, CDR2, and CDR3 regions consist of SEQ ID NO:433, 456, and 401, respectively. In some embodiments, the antibodycomprises the same VH CDRs as G9.2-12.

In some embodiments, the anti-Galectin-9 antibody or binding portionthereof comprises heavy and light chain variable regions, wherein theheavy chain variable region CDR1, CDR2, and CDR3 regions comprise SEQ IDNO: 434, 362, and 402, respectively. In some embodiments, the heavychain variable region CDR1, CDR2, and CDR3 regions consist of SEQ ID NO:434, 362, and 402, respectively. In some embodiments, the antibodycomprises the same VH CDRs as G9.2-13.

In some embodiments, the anti-Galectin-9 antibody or binding portionthereof comprises heavy and light chain variable regions, wherein theheavy chain variable region CDR1, CDR2, and CDR3 regions comprise SEQ IDNO: 361, 457, and 403, respectively. In some embodiments, the heavychain variable region CDR1, CDR2, and CDR3 regions consist of SEQ ID NO:361, 457, and 403, respectively. In some embodiments, the antibodycomprises the same VH CDRs as G9.2-14.

In some embodiments, the anti-Galectin-9 antibody or binding portionthereof comprises heavy and light chain variable regions, wherein theheavy chain variable region CDR1, CDR2, and CDR3 regions comprise SEQ IDNO: 361, 458, and 404, respectively. In some embodiments, the heavychain variable region CDR1, CDR2, and CDR3 regions consist of SEQ ID NO:361, 458, and 404, respectively. In some embodiments, the antibodycomprises the same VH CDRs as G9.2-15.

In some embodiments, the anti-Galectin-9 antibody or binding portionthereof comprises heavy and light chain variable regions, wherein theheavy chain variable region CDR1, CDR2, and CDR3 regions comprise SEQ IDNO: 361, 459, and 405, respectively. In some embodiments, the heavychain variable region CDR1, CDR2, and CDR3 regions consist of SEQ ID NO:361, 459, and 405, respectively. In some embodiments, the antibodycomprises the same VH CDRs as G9.2-16.

In some embodiments, the anti-Galectin-9 antibody or binding portionthereof comprises heavy and light chain variable regions, wherein theheavy chain variable region CDR1, CDR2, and CDR3 regions comprise SEQ IDNO: 361, 388, and 406, respectively. In some embodiments, the heavychain variable region CDR1, CDR2, and CDR3 regions consist of SEQ ID NO:361, 388, and 406. In some embodiments, the antibody comprises the sameVH CDRs as G9.2-17.

In some embodiments, the anti-Galectin-9 antibody or binding portionthereof comprises heavy and light chain variable regions, wherein theheavy chain variable region CDR1, CDR2, and CDR3 regions comprise SEQ IDNO: 361, 388, and 407, respectively. In some embodiments, the heavychain variable region CDR1, CDR2, and CDR3 regions consist of SEQ ID NO:361, 388, and 407, respectively. In some embodiments, the antibodycomprises the same VH CDRs as G9.2-17mut6.

In some embodiments, the anti-Galectin-9 antibody or binding portionthereof comprises heavy and light chain variable regions, wherein theheavy chain variable region CDR1, CDR2, and CDR3 regions comprise SEQ IDNO: 430, 363, and 408, respectively. In some embodiments, the heavychain variable region CDR1, CDR2, and CDR3 regions consist of SEQ ID NO:430, 363, and 408, respectively. In some embodiments, the antibodycomprises the same VH CDRs as G9.2-18.

In some embodiments, the anti-Galectin-9 antibody or binding portionthereof comprises heavy and light chain variable regions, wherein theheavy chain variable region CDR1, CDR2, and CDR3 regions comprise SEQ IDNO: 430, 460, and 409, respectively. In some embodiments, the heavychain variable region CDR1, CDR2, and CDR3 regions consist of SEQ ID NO:430, 460, and 409, respectively. In some embodiments, the antibodycomprises the same VH CDRs as G9.2-19.

In some embodiments, the anti-Galectin-9 antibody or binding portionthereof comprises heavy and light chain variable regions, wherein theheavy chain variable region CDR1, CDR2, and CDR3 regions comprise SEQ IDNO: 429, 461, and 410, respectively. In some embodiments, the heavychain variable region CDR1, CDR2, and CDR3 regions consist of SEQ ID NO:429, 461, and 410, respectively. In some embodiments, the antibodycomprises the same VH CDRs as G9.2-20.

In some embodiments, the anti-Galectin-9 antibody or binding portionthereof comprises heavy and light chain variable regions, wherein theheavy chain variable region CDR1, CDR2, and CDR3 regions comprise SEQ IDNO: 429, 462, and 411, respectively. In some embodiments, the heavychain variable region CDR1, CDR2, and CDR3 regions consist of SEQ ID NO:429, 462, and 411, respectively. In some embodiments, the antibodycomprises the same VH CDRs as G9.2-21.

In some embodiments, the anti-Galectin-9 antibody or binding portionthereof comprises heavy and light chain variable regions, wherein theheavy chain variable region CDR1, CDR2, and CDR3 regions comprise SEQ IDNO: 428, 463, and 412, respectively. In some embodiments, the heavychain variable region CDR1, CDR2, and CDR3 regions consist of SEQ ID NO:428, 463, and 412, respectively. In some embodiments, the antibodycomprises the same VH CDRs as G9.2-22.

In some embodiments, the anti-Galectin-9 antibody or binding portionthereof comprises heavy and light chain variable regions, wherein theheavy chain variable region CDR1, CDR2, and CDR3 regions comprise SEQ IDNO: 428, 464, and 413, respectively. In some embodiments, the heavychain variable region CDR1, CDR2, and CDR3 regions consist of SEQ ID NO:428, 464, and 413, respectively. In some embodiments, the antibodycomprises the same VH CDRs as G9.2-23.

In some embodiments, the anti-Galectin-9 antibody or binding portionthereof comprises heavy and light chain variable regions, wherein theheavy chain variable region CDR1, CDR2, and CDR3 regions comprise SEQ IDNO: 428, 465, and 414, respectively. In some embodiments, the heavychain variable region CDR1, CDR2, and CDR3 regions consist of SEQ ID NO:428, 465, and 414, respectively. In some embodiments, the antibodycomprises the same VH CDRs as G9.2-24.

In some embodiments, the anti-Galectin-9 antibody or binding portionthereof comprises heavy and light chain variable regions, wherein theheavy chain variable region CDR1, CDR2, and CDR3 regions comprise SEQ IDNO: 427, 466, and 415, respectively. In some embodiments, the heavychain variable region CDR1, CDR2, and CDR3 regions consist of SEQ ID NO:427, 466, and 415. In some embodiments, the antibody comprises the sameVH CDRs as G9.2-25.

In some embodiments, the anti-Galectin-9 antibody or binding portionthereof comprises heavy and light chain variable regions, wherein theheavy chain variable region CDR1, CDR2, and CDR3 regions comprise SEQ IDNO: 361, 389, and 416, respectively. In some embodiments, the heavychain variable region CDR1, CDR2, and CDR3 regions consist of SEQ ID NO:361, 389, and 416, respectively. In some embodiments, the antibodycomprises the same VH CDRs as G9.2-26.

In some embodiments, the anti-Galectin-9 antibody or binding portionthereof comprises heavy and light chain variable regions, wherein theheavy chain variable region CDR1, CDR2, and CDR3 regions comprise SEQ IDNO: 361, 388, and 417, respectively. In some embodiments, the heavychain variable region CDR1, CDR2, and CDR3 regions consist of SEQ ID NO:361, 388, and 417, respectively. In some embodiments, the antibodycomprises the same VH CDRs as G9.2-low affinity binder.

Clone 9.2-Derived Heavy and Light Chain Variable Regions

In some embodiments, the anti-Galectin-9 antibody or binding portionthereof comprises heavy and light chain variable regions, wherein thelight chain variable region CDR1, CDR2, and CDR3 regions comprise SEQ IDNO: 328, 329, and 341, respectively, and the heavy chain variable regionCDR1, CDR2, and CDR3 comprise SEQ ID NO: 424, 446, and 390 respectively.In some embodiments, the light and heavy chain variable region CDR1,CDR2, and CDR3 regions consist of SEQ ID NO: 328, 329, and 341, and SEQID NO: 424, 446, and 390. In one specific embodiment, the antibodycomprises the same VL and VH CDRs as G9.2-1.

In some embodiments, the anti-Galectin-9 antibody or binding portionthereof comprises heavy and light chain variable regions, wherein thelight chain variable region CDR1, CDR2, and CDR3 regions comprise SEQ IDNO: 328, 329, and 333, respectively, and the heavy chain variable regionCDR1, CDR2, and CDR3 comprise SEQ ID NO: 431, 447, and 391,respectively. In some embodiments, the light and heavy chain variableregion CDR1, CDR2, and CDR3 regions consist of SEQ ID NO: 328, 329, and333, and SEQ ID NO: 431, 447, and 391. In one specific embodiment, theantibody comprises the same VL and VH CDRs as G9.2-2.

In some embodiments, the anti-Galectin-9 antibody or binding portionthereof comprises heavy and light chain variable regions, wherein thelight chain variable region CDR1, CDR2, and CDR3 regions comprise SEQ IDNO: 328, 329, and 333, respectively, and the heavy chain variable regionCDR1, CDR2, and CDR3 comprise SEQ ID NO: 431, 448, and 392,respectively. In some embodiments, the light and heavy chain variableregion CDR1, CDR2, and CDR3 regions consist of SEQ ID NO: 328, 329, and333, and SEQ ID NO: 431, 448, and 392. In one specific embodiment, theantibody comprises the same VL and VH CDRs as G9.2-3.

In some embodiments, the anti-Galectin-9 antibody or binding portionthereof comprises heavy and light chain variable regions, wherein thelight chain variable region CDR1, CDR2, and CDR3 regions comprise SEQ IDNO: 328, 329, and 342, respectively, and the heavy chain variable regionCDR1, CDR2, and CDR3 comprise SEQ ID NO: 431, 449, and 393,respectively. In some embodiments, the light and heavy chain variableregion CDR1, CDR2, and CDR3 regions consist of SEQ ID NO: 328, 329, and342, and SEQ ID NO: 431, 449, and 393. In one specific embodiment, theantibody comprises the same VL and VH CDRs as G9.2-4.

In some embodiments, the anti-Galectin-9 antibody or binding portionthereof comprises heavy and light chain variable regions, wherein thelight chain variable region CDR1, CDR2, and CDR3 regions comprise SEQ IDNO: 328, 329, and 343, respectively, and the heavy chain variable regionCDR1, CDR2, and CDR3 comprise SEQ ID NO: 431, 450, and 394,respectively. In some embodiments, the light and heavy chain variableregion CDR1, CDR2, and CDR3 regions consist of SEQ ID NO: 328, 329, and343, and SEQ ID NO: 431, 450, and 394. In one specific embodiment, theantibody comprises the same VL and VH CDRs as G9.2-5.

In some embodiments, the anti-Galectin-9 antibody or binding portionthereof comprises heavy and light chain variable regions, wherein thelight chain variable region CDR1, CDR2, and CDR3 regions comprise SEQ IDNO: 328, 329, and 344, respectively, and the heavy chain variable regionCDR1, CDR2, and CDR3 comprise SEQ ID NO: 431, 451, and 395,respectively. In some embodiments, the light and heavy chain variableregion CDR1, CDR2, and CDR3 regions consist of SEQ ID NO: 328, 329, and344, and SEQ ID NO: 431, 451, and 395. In one specific embodiment, theantibody comprises the same VL and VH CDRs as G9.2-6.

In some embodiments, the anti-Galectin-9 antibody or binding portionthereof comprises heavy and light chain variable regions, wherein thelight chain variable region CDR1, CDR2, and CDR3 regions comprise SEQ IDNO: 328, 329, and 345, respectively, and the heavy chain variable regionCDR1, CDR2, and CDR3 comprise SEQ ID NO: 425, 452, and 396,respectively. In some embodiments, the light and heavy chain variableregion CDR1, CDR2, and CDR3 regions consist of SEQ ID NO: 328, 329, and345, and SEQ ID NO: 425, 452, and 396. In one specific embodiment, theantibody comprises the same VL and VH CDRs as G9.2-7.

In some embodiments, the anti-Galectin-9 antibody or binding portionthereof comprises heavy and light chain variable regions, wherein thelight chain variable region CDR1, CDR2, and CDR3 regions comprise SEQ IDNO: 328, 329, and 346, respectively, and the heavy chain variable regionCDR1, CDR2, and CDR3 comprise SEQ ID NO: 245, 453, and 397,respectively. In some embodiments, the light and heavy chain variableregion CDR1, CDR2, and CDR3 regions consist of SEQ ID NO: 328, 329, and346, and SEQ ID NO: 245, 453, and 397. In one specific embodiment, theantibody comprises the same VL and VH CDRs as G9.2-8.

In some embodiments, the anti-Galectin-9 antibody or binding portionthereof comprises heavy and light chain variable regions, wherein thelight chain variable region CDR1, CDR2, and CDR3 regions comprise SEQ IDNO: 328, 329, and 347, respectively, and the heavy chain variable regionCDR1, CDR2, and CDR3 comprise SEQ ID NO: 426, 454, and 398,respectively. In some embodiments, the light and heavy chain variableregion CDR1, CDR2, and CDR3 regions consist of SEQ ID NO: 328, 329, and347, and SEQ ID NO: 426, 454, and 398. In one specific embodiment, theantibody comprises the same VL and VH CDRs as G9.2-9.

In some embodiments, the anti-Galectin-9 antibody or binding portionthereof comprises heavy and light chain variable regions, wherein thelight chain variable region CDR1, CDR2, and CDR3 regions comprise SEQ IDNO: 328, 329, and 348, respectively, and the heavy chain variable regionCDR1, CDR2, and CDR3 comprise SEQ ID NO: 426, 387, and 399,respectively. In some embodiments, the light and heavy chain variableregion CDR1, CDR2, and CDR3 regions consist of SEQ ID NO: 328, 329, and348, and SEQ ID NO: 426, 387, and 399. In one specific embodiment, theantibody comprises the same VL and VH CDRs as G9.2-10.

In some embodiments, the anti-Galectin-9 antibody or binding portionthereof comprises heavy and light chain variable regions, wherein thelight chain variable region CDR1, CDR2, and CDR3 regions comprise SEQ IDNO: 328, 329, and 349, respectively, and the heavy chain variable regionCDR1, CDR2, and CDR3 comprise SEQ ID NO: 432, 455, and 400,respectively. In some embodiments, the light and heavy chain variableregion CDR1, CDR2, and CDR3 regions consist of SEQ ID NO: 328, 329, and349, and SEQ ID NO: 432, 455, and 400. In one specific embodiment, theantibody comprises the same VL and VH CDRs as G9.2-11.

In some embodiments, the anti-Galectin-9 antibody or binding portionthereof comprises heavy and light chain variable regions, wherein thelight chain variable region CDR1, CDR2, and CDR3 regions comprise SEQ IDNO: 328, 329, and 350, respectively, and the heavy chain variable regionCDR1, CDR2, and CDR3 comprise SEQ ID NO: 433, 456, and 401,respectively. In some embodiments, the light and heavy chain variableregion CDR1, CDR2, and CDR3 regions consist of SEQ ID NO: 328, 329, and350, and SEQ ID NO: 433, 456, and 401. In one specific embodiment, theantibody comprises the same VL and VH CDRs as G9.2-12.

In some embodiments, the anti-Galectin-9 antibody or binding portionthereof comprises heavy and light chain variable regions, wherein thelight chain variable region CDR1, CDR2, and CDR3 regions comprise SEQ IDNO: 328, 329, and 341, respectively, and the heavy chain variable regionCDR1, CDR2, and CDR3 comprise SEQ ID NO: 434, 362, and 402,respectively. In some embodiments, the light and heavy chain variableregion CDR1, CDR2, and CDR3 regions consist of SEQ ID NO: 328, 329, and341, and SEQ ID NO: 434, 362, and 402. In one specific embodiment, theantibody comprises the same VL and VH CDRs as G9.2-13.

In some embodiments, the anti-Galectin-9 antibody or binding portionthereof comprises heavy and light chain variable regions, wherein thelight chain variable region CDR1, CDR2, and CDR3 regions comprise SEQ IDNO: 328, 329, and 333, respectively, and the heavy chain variable regionCDR1, CDR2, and CDR3 comprise SEQ ID NO: 361, 457, and 403,respectively. In some embodiments, the light and heavy chain variableregion CDR1, CDR2, and CDR3 regions consist of SEQ ID NO: 328, 329, and333, and SEQ ID NO: 361, 457, and 403. In one specific embodiment, theantibody comprises the same VL and VH CDRs as G9.2-14.

In some embodiments, the anti-Galectin-9 antibody or binding portionthereof comprises heavy and light chain variable regions, wherein thelight chain variable region CDR1, CDR2, and CDR3 regions comprise SEQ IDNO: 328, 329, and 343, respectively, and the heavy chain variable regionCDR1, CDR2, and CDR3 comprise SEQ ID NO: 361, 458, and 404,respectively. In some embodiments, the light and heavy chain variableregion CDR1, CDR2, and CDR3 regions consist of SEQ ID NO: 328, 329, and343, and SEQ ID NO: 361, 458, and 404. In one specific embodiment, theantibody comprises the same VL and VH CDRs as G9.2-15.

In some embodiments, the anti-Galectin-9 antibody or binding portionthereof comprises heavy and light chain variable regions, wherein thelight chain variable region CDR1, CDR2, and CDR3 regions comprise SEQ IDNO: 328, 329, and 333, respectively, and the heavy chain variable regionCDR1, CDR2, and CDR3 comprise SEQ ID NO: 361, 459, and 405,respectively. In some embodiments, the light and heavy chain variableregion CDR1, CDR2, and CDR3 regions consist of SEQ ID NO: 328, 329, and333, and SEQ ID NO: 361, 459, and 405. In one specific embodiment, theantibody comprises the same VL and VH CDRs as G9.2-16.

In some embodiments, the anti-Galectin-9 antibody or binding portionthereof comprises heavy and light chain variable regions, wherein thelight chain variable region CDR1, CDR2, and CDR3 regions comprise SEQ IDNO: 328, 329, and 352, respectively, and the heavy chain variable regionCDR1, CDR2, and CDR3 comprise SEQ ID NO: 361, 388, and 406,respectively. In some embodiments, the light and heavy chain variableregion CDR1, CDR2, and CDR3 regions consist of SEQ ID NO: 328, 329, and352, and SEQ ID NO: 361, 388, and 406. In one specific embodiment, theantibody comprises the same VL and VH CDRs as G9.2-17.

In some embodiments, the anti-Galectin-9 antibody or binding portionthereof comprises heavy and light chain variable regions, wherein thelight chain variable region CDR1, CDR2, and CDR3 regions comprise SEQ IDNO: 328, 329, and 352, respectively, and the heavy chain variable regionCDR1, CDR2, and CDR3 comprise SEQ ID NO: 361, 388, and 404,respectively. In some embodiments, the light and heavy chain variableregion CDR1, CDR2, and CDR3 regions consist of SEQ ID NO: 328, 329, and352, and SEQ ID NO: 361, 388, and 404. In one specific embodiment, theantibody comprises the same VL and VH CDRs as G9.2-17mut6.

In some embodiments, the anti-Galectin-9 antibody or binding portionthereof comprises heavy and light chain variable regions, wherein thelight chain variable region CDR1, CDR2, and CDR3 regions comprise SEQ IDNO: 328, 329, and 333, respectively, and the heavy chain variable regionCDR1, CDR2, and CDR3 comprise SEQ ID NO: 430, 363, and 408,respectively. In some embodiments, the light and heavy chain variableregion CDR1, CDR2, and CDR3 regions consist of SEQ ID NO: 328, 329, and333, and SEQ ID NO: 430, 363, and 408. In one specific embodiment, theantibody comprises the same VL and VH CDRs as G9.2-18.

In some embodiments, the anti-Galectin-9 antibody or binding portionthereof comprises heavy and light chain variable regions, wherein thelight chain variable region CDR1, CDR2, and CDR3 regions comprise SEQ IDNO: 328, 329, and 354, respectively, and the heavy chain variable regionCDR1, CDR2, and CDR3 comprise SEQ ID NO: 430, 460, and 409,respectively. In some embodiments, the light and heavy chain variableregion CDR1, CDR2, and CDR3 regions consist of SEQ ID NO: 328, 329, and354, and SEQ ID NO: 430, 460, and 409. In one specific embodiment, theantibody comprises the same VL and VH CDRs as G9.2-19.

In some embodiments, the anti-Galectin-9 antibody or binding portionthereof comprises heavy and light chain variable regions, wherein thelight chain variable region CDR1, CDR2, and CDR3 regions comprise SEQ IDNO: 328, 329, and 352, respectively, and the heavy chain variable regionCDR1, CDR2, and CDR3 comprise SEQ ID NO: 429, 461, and 410,respectively. In some embodiments, the light and heavy chain variableregion CDR1, CDR2, and CDR3 regions consist of SEQ ID NO: 328, 329, and352, and SEQ ID NO: 429, 461, and 410. In one specific embodiment, theantibody comprises the same VL and VH CDRs as G9.2-20.

In some embodiments, the anti-Galectin-9 antibody or binding portionthereof comprises heavy and light chain variable regions, wherein thelight chain variable region CDR1, CDR2, and CDR3 regions comprise SEQ IDNO: 328, 329, and 355, respectively, and the heavy chain variable regionCDR1, CDR2, and CDR3 comprise SEQ ID NO: 429, 462, and 411,respectively. In some embodiments, the light and heavy chain variableregion CDR1, CDR2, and CDR3 regions consist of SEQ ID NO: 328, 329, and355, and SEQ ID NO: 429, 462, and 411. In one specific embodiment, theantibody comprises the same VL and VH CDRs as G9.2-21.

In some embodiments, the anti-Galectin-9 antibody or binding portionthereof comprises heavy and light chain variable regions, wherein thelight chain variable region CDR1, CDR2, and CDR3 regions comprise SEQ IDNO: 328, 329, and 356, respectively, and the heavy chain variable regionCDR1, CDR2, and CDR3 comprise SEQ ID NO: 428, 463, and 412,respectively. In some embodiments, the light and heavy chain variableregion CDR1, CDR2, and CDR3 regions consist of SEQ ID NO: 328, 329, and356, and SEQ ID NO: 428, 463, and 412. In one specific embodiment, theantibody comprises the same VL and VH CDRs as G9.2-22.

In some embodiments, the anti-Galectin-9 antibody or binding portionthereof comprises heavy and light chain variable regions, wherein thelight chain variable region CDR1, CDR2, and CDR3 regions comprise SEQ IDNO: 328, 329, and 357, respectively, and the heavy chain variable regionCDR1, CDR2, and CDR3 comprise SEQ ID NO: 428, 464, and 413,respectively. In some embodiments, the light and heavy chain variableregion CDR1, CDR2, and CDR3 regions consist of SEQ ID NO: 328, 329, and357, and SEQ ID NO: 428, 464, and 413. In one specific embodiment, theantibody comprises the same VL and VH CDRs as G9.2-23.

In some embodiments, the anti-Galectin-9 antibody or binding portionthereof comprises heavy and light chain variable regions, wherein thelight chain variable region CDR1, CDR2, and CDR3 regions comprise SEQ IDNO: 328, 329, and 358, respectively, and the heavy chain variable regionCDR1, CDR2, and CDR3 comprise SEQ ID NO: 428, 465, and 414,respectively. In some embodiments, the light and heavy chain variableregion CDR1, CDR2, and CDR3 regions consist of SEQ ID NO: 328, 329, and358, and SEQ ID NO: 428, 465, and 414. In one specific embodiment, theantibody comprises the same VL and VH CDRs as G9.2-24.

In some embodiments, the anti-Galectin-9 antibody or binding portionthereof comprises heavy and light chain variable regions, wherein thelight chain variable region CDR1, CDR2, and CDR3 regions comprise SEQ IDNO: 328, 329, and 359, respectively, and the heavy chain variable regionCDR1, CDR2, and CDR3 comprise SEQ ID NO: 427, 466, and 415,respectively. In some embodiments, the light and heavy chain variableregion CDR1, CDR2, and CDR3 regions consist of SEQ ID NO: 328, 329, and359, and SEQ ID NO: 427, 466, and 415. In one specific embodiment, theantibody comprises the same VL and VH CDRs as G9.2-25.

In some embodiments, the anti-Galectin-9 antibody or binding portionthereof comprises heavy and light chain variable regions, wherein thelight chain variable region CDR1, CDR2, and CDR3 regions comprise SEQ IDNO: 328, 329, and 360, respectively, and the heavy chain variable regionCDR1, CDR2, and CDR3 comprise SEQ ID NO: 361, 389, and 416,respectively. In some embodiments, the light and heavy chain variableregion CDR1, CDR2, and CDR3 regions consist of SEQ ID NO: 328, 329, and360, and SEQ ID NO: 361, 389, and 416. In one specific embodiment, theantibody comprises the same VL and VH CDRs as G9.2-26.

In some embodiments, the anti-Galectin-9 antibody or binding portionthereof comprises heavy and light chain variable regions, wherein thelight chain variable region CDR1, CDR2, and CDR3 regions comprise SEQ IDNO: 328, 329, and 352, respectively, and the heavy chain variable regionCDR1, CDR2, and CDR3 comprise SEQ ID NO: 361, 388, and 417,respectively. In some embodiments, the light and heavy chain variableregion CDR1, CDR2, and CDR3 regions consist of SEQ ID NO: 328, 329, and352, and SEQ ID NO: 361, 388, and 417. In one specific embodiment, theantibody comprises the same VL and VH CDRs as G9.2-low affinity binder.

Sequence Identity

In some embodiments, the anti-Galectin-9 antibody (e.g., specific toCRD1 and/or CRD2) may comprise light chain CDRs that have at least 80%(e.g., 85%, 90%, 95%, 96%, 97%, 98%, 99% and any increment therein)sequence identity, individually or collectively, as compared with thecorresponding V_(L) CDRs of an antibody or antigen binding portionthereof selected from G9.2-1, G9.2-2, G9.2-3, G9.2-4, G9.2-5, G9.2-6,G9.2-7, G9.2-8, G9.2-9, G9.2-10, G9.2-11, G9.2-12, G9.2-13, G9.2-14,G9.2-15, G9.2-16, G9.2-17, G9.2-17mut6, G9.2-18, G9.2-19, G9.2-20,G9.2-21, G9.2-22, G9.2-23, G9.2-24, G9.2-25, G9.2-26, and G9.2-lowaffinity binder. In some embodiments, the anti-Galectin-9 antibody(e.g., specific to CRD1 and/or CRD2) may comprise heavy chain CDRs thathave at least 80% (e.g., 85%, 90%, 95%, 96%, 97%, 98%, 99% and anyincrement therein) sequence identity, individually or collectively, ascompared with the corresponding V_(H) CDRs of an antibody or antigenbinding portion thereof selected from G9.2-1, G9.2-2, G9.2-3, G9.2-4,G9.2-5, G9.2-6, G9.2-7, G9.2-8, G9.2-9, G9.2-10, G9.2-11, G9.2-12,G9.2-13, G9.2-14, G9.2-15, G9.2-16, G9.2-17, G9.2-17mut6, G9.2-18,G9.2-19, G9.2-20, G9.2-21, G9.2-22, G9.2-23, G9.2-24, G9.2-25, G9.2-26,and G9.2-low affinity binder.

In some embodiments, the anti-Galectin-9 antibody (e.g., specific toCRD1 and/or CRD2) may comprise light chain CDRs and heavy chain CDRsthat have at least 80% (e.g., 85%, 90%, 95%, 96%, 97%, 98%, 99% and anyincrement therein) sequence identity, individually or collectively, ascompared with the corresponding V_(L) CDRs and V_(H) CDRs of an antibodyor antigen binding portion thereof selected from G9.2-1, G9.2-2, G9.2-3,G9.2-4, G9.2-5, G9.2-6, G9.2-7, G9.2-8, G9.2-9, G9.2-10, G9.2-11,G9.2-12, G9.2-13, G9.2-14, G9.2-15, G9.2-16, G9.2-17, G9.2-17mut6,G9.2-18, G9.2-19, G9.2-20, G9.2-21, G9.2-22, G9.2-23, G9.2-24, G9.2-25,G9.2-26, and G9.2-low affinity binder.

In some embodiments, the anti-Galectin-9 antibody (e.g., specific toCRD1 and/or CRD2) may comprise a VL CDR1 amino acid sequence that has atleast 80% (e.g., 85%, 90%, 95%, 96%, 97%, 98%, 99% and any incrementtherein) sequence identity to the VL CDR1 amino acid sequence set forthin SEQ ID NO: 328. In some embodiments, the anti-Galectin-9 antibody(e.g., specific to CRD1 and/or CRD2) may comprise a VL CDR2 amino acidsequence that has at least 80% (e.g., 85%, 90%, 95%, 96%, 97%, 98%, 99%and any increment therein) sequence identity to the VL CDR2 amino acidsequence set forth in SEQ ID NO: 329. In some embodiments, theanti-Galectin-9 antibody (e.g., specific to CRD1 and/or CRD2) maycomprise a VL CDR3 amino acid sequence that has at least 80% (e.g., 85%,90%, 95%, 96%, 97%, 98%, 99% and any increment therein) sequenceidentity to a VL CDR3 amino acid sequence selected from SEQ ID NO:341-360.

In some embodiments, the anti-Galectin-9 antibody (e.g., specific toCRD1 and/or CRD2) may comprise a VH CDR1 amino acid sequence that has atleast 80% (e.g., 85%, 90%, 95%, 96%, 97%, 98%, 99% and any incrementtherein) sequence identity to the VH CDR1 amino acid sequence set forthin SEQ ID NO: 361, 424-434. In some embodiments, the anti-Galectin-9antibody (e.g., specific to CRD1 and/or CRD2) may comprise a VH CDR2amino acid sequence that has at least 80% (e.g., 85%, 90%, 95%, 96%,97%, 98%, 99% and any increment therein) sequence identity to a VH CDR2amino acid sequence selected from SEQ ID NO: 362, 363, 387-389 and446-466. In some embodiments, the anti-Galectin-9 antibody (e.g.,specific to CRD1 and/or CRD2) may comprise a VH CDR3 amino acid sequencethat has at least 80% (e.g., 85%, 90%, 95%, 96%, 97%, 98%, 99% and anyincrement therein) sequence identity to a VH CDR3 amino acid sequenceselected from SEQ ID NO: 390-417.

Accordingly, in some embodiments, anti-Galectin-9 antibodies or antigenbinding portions thereof comprise (a) VL CDR1 amino acid sequence thathas at least 80% (e.g., 85%, 90%, 95%, 96%, 97%, 98%, 99% and anyincrement therein) sequence identity to the VL CDR1 amino acid sequenceset forth in SEQ ID NO: 328; (b) VL CDR2 amino acid sequence that has atleast 80% (e.g., 85%, 90%, 95%, 96%, 97%, 98%, 99% and any incrementtherein) sequence identity to the VL CDR2 amino acid sequence set forthin SEQ ID NO: 329; (c) VL CDR3 amino acid sequence that has at least 80%(e.g., 85%, 90%, 95%, 96%, 97%, 98%, 99% and any increment therein)sequence identity to the VL CDR3 amino acid sequence selected from SEQID NO: 341-360; (d) VH CDR1 amino acid sequence that has at least 80%(e.g., 85%, 90%, 95%, 96%, 97%, 98%, 99% and any increment therein)sequence identity to the VH CDR1 amino acid sequence set forth in SEQ IDNO: 361, 424-434; (d) VH CDR2 amino acid sequence that has at least 80%(e.g., 85%, 90%, 95%, 96%, 97%, 98%, 99% and any increment therein)sequence identity to a VH CDR2 amino acid sequence selected from SEQ IDNO: 362, 363, 387-389 and 446-466; (e) VH CDR3 amino acid sequence thathas at least 80% (e.g., 85%, 90%, 95%, 96%, 97%, 98%, 99% and anyincrement therein) sequence identity to a VH CDR3 amino acid sequenceselected from SEQ ID NO: 390-417.

In some embodiments, the anti-Galectin-9 antibody or binding portionthereof comprises heavy and light chain variable regions, wherein thelight chain variable region CDR1, CDR2, and CDR3 amino acid sequenceshave at least 80% (e.g., 85%, 90%, 95%, 96%, 97%, 98%, 99% and anyincrement therein) sequence identity to the light chain variable regionCDR1, CDR2, and CDR3 amino acid sequences set forth in SEQ ID NO: 328,329, and 352, respectively. In some embodiments, the antibody VL CDR1,CDR2, and CDR3 amino acid sequences have at least 80% (e.g., 85%, 90%,95%, 96%, 97%, 98%, 99% and any increment therein) sequence identity tothe VL CDR1, CDR2, and CDR3 amino acid sequences of G9.2-17. In someembodiments, the anti-Galectin-9 antibody or binding portion thereofcomprises heavy and light chain variable regions, wherein the heavychain variable region CDR1, CDR2, and CDR3 amino acid sequences have atleast 80% (e.g., 85%, 90%, 95%, 96%, 97%, 98%, 99% and any incrementtherein) sequence identity to the heavy chain variable region CDR1,CDR2, and CDR3 amino acid sequences set forth in SEQ ID NO: 361, 388,and 406, respectively. In some embodiments, the antibody VH CDR1, CDR2,and CDR3 amino acid sequences have at least 80% (e.g., 85%, 90%, 95%,96%, 97%, 98%, 99% and any increment therein) sequence identity to theVH CDR1, CDR2, and CDR3 amino acid sequences of G9.2-17. In someembodiments, the anti-Galectin-9 antibody or binding portion thereofcomprises heavy and light chain variable regions, wherein the lightchain variable region CDR1, CDR2, and CDR3 amino acid sequences have atleast 80% (e.g., 85%, 90%, 95%, 96%, 97%, 98%, 99% and any incrementtherein) sequence identity to the light chain variable region CDR1,CDR2, and CDR3 amino acid sequences set forth in comprise SEQ ID NO:328, 329, and 352, respectively, and the heavy chain variable regionCDR1, CDR2, and CDR3 amino acid sequences have at least 80% (e.g., 85%,90%, 95%, 96%, 97%, 98%, 99% and any increment therein) sequenceidentity to the heavy chain variable region CDR1, CDR2, and CDR3 aminoacid sequences set forth in SEQ ID NO: 361, 388, and 406, respectively.In one specific embodiment, the antibody VL CDR1, CDR2, and CDR3 and VHCDR1, CDR2, and CDR3 amino acid sequences have at least 80% (e.g., 85%,90%, 95%, 96%, 97%, 98%, 99% and any increment therein) sequenceidentity to the VL CDR1, CDR2, and CDR3 and VH CDR1, CDR2, and CDR3amino acid sequences of G9.2-17.

Epitopes and Constant Regions

In some embodiments, the anti-Galectin-9 antibodies described hereinbind to the same epitope as any of the exemplary antibodies describedherein (e.g., antibody comprising any of SEQ ID NO: 7-87 or the CDRsthereof) or competes against the exemplary antibody from binding to theGalectin-9 antigen. An “epitope” refers to the site on a target antigenthat is recognized and bound by an antibody. The site can be entirelycomposed of amino acid components, entirely composed of chemicalmodifications of amino acids of the protein (e.g., glycosyl moieties),or composed of combinations thereof. Overlapping epitopes include atleast one common amino acid residue. An epitope can be linear, which istypically 6-15 amino acids in length. Alternatively, the epitope can beconformational. The epitope to which an antibody binds can be determinedby routine technology, for example, the epitope mapping method (see,e.g., descriptions below). An antibody that binds the same epitope as anexemplary antibody described herein may bind to exactly the same epitopeor a substantially overlapping epitope (e.g., containing less than 3non-overlapping amino acid residue, less than 2 non-overlapping aminoacid residues, or only 1 non-overlapping amino acid residue) as theexemplary antibody. Whether two antibodies compete against each otherfrom binding to the cognate antigen can be determined by a competitionassay, which is well known in the art.

In some embodiments, the anti-Galectin-9 antibody may bind to an epitopeat least a segment of which is in CRD1 of a galectin-9 protein (e.g., ahuman galectin-9 or a mouse galectin-9). In some embodiments, theantibody may bind an epitope which is entirely within the CRD1 of theGalectin-9 protein. In some embodiments, the antibody may bind anepitope which is partially within the CRD1 of the Galectin-9 protein. Insome embodiments, the epitope to which the anti-Galectin antibody bindsis a linear epitope. In some embodiments, the epitope to which theanti-Galectin antibody binds is a conformational epitope.

In some embodiments, the anti-Galectin-9 antibody may bind an epitope atleast a segment of which is in CRD2 of a Galectin-9 protein (e.g., ahuman galectin-9 or a mouse galectin-9). In some embodiments, theanti-Galectin-9 antibody may bind an epitope which is entirely withinthe CRD2 of the Galectin-9 protein. In some specific embodiments inwhich the anti-Galectin-9 antibody binds an epitope partially orentirely within CDR2, the antibody binds an epitope comprising at leastresidue W309. In some specific embodiments, in which the anti-Galectin-9antibody binds an epitope partially or entirely within CDR2, the epitopeto which the anti-Galectin-9 antibody binds does not contain one or moreof R253, R271, Y330, R334, R341, and Y236 of SEQ ID NO:1. In someembodiments, the epitope to which the anti-Galectin antibody binds is alinear epitope encompassing residue W309. In some embodiments, theepitope to which the anti-Galectin antibody binds is a conformationalepitope comprising W309.

In some examples, the anti-Galectin-9 antibody comprises the same V_(H)and/or V_(L) CDRs as an exemplary antibody described herein. Twoantibodies having the same V_(H) and/or V_(L) CDRs means that their CDRsare identical when determined by the same approach (e.g., the Kabatapproach or the Chothia approach as known in the art). Suchanti-Galectin-9 antibodies may have the same V_(H), the same V_(L), orboth as compared to an exemplary antibody described herein.

Two heavy chain variable regions (or two light chain variable regions)having the same CDRs means that the CDRs in the two heavy chain variableregions (or light chain variable regions) as determined by the samenumbering scheme are identical. Exemplary numbering schemes fordetermining antibody CDRs include the “Kabat” numbering scheme (Kabat etal. (1991), 5th Ed. Public Health Service, National Institutes ofHealth, Bethesda, Md.), the “Chothia” numbering scheme (Al-Lazikani etal., (1997) JMB 273, 927-948), the “Contact” numbering scheme (MacCallumet al., J. Mol. Biol. 262:732-745 (1996)), the “IMGT” numbering scheme(Lefranc M P et al., Dev Comp Immunol, 2003 January; 27(1):55-77), andthe “AHo” numbering scheme (Honegger A and Pluckthun A, J Mol Biol, 2001Jun. 8; 309(3):657-70). As known to those skilled in the art, the CDRregions of the exemplary anti-pKal and anti-FXII antibodies identifiedherein are determined by the “Chothia” numbering scheme, which is usedas an example.

Also within the scope of the present disclosure are functional variantsof any of the exemplary anti-Galectin-9 antibodies as disclosed herein.Such functional variants are substantially similar to the exemplaryantibody, both structurally and functionally. A functional variantcomprises substantially the same V_(H) and V_(L) CDRs as the exemplaryantibody. For example, it may comprise only up to 5 (e.g., 4, 3, 2,or 1) amino acid residue variations in the total CDR regions of theantibody and binds the same epitope of Galectin-9 with substantiallysimilar affinity (e.g., having a K_(D) value in the same order).Alternatively or in addition, the amino acid residue variations areconservative amino acid residue substitutions. As used herein, a“conservative amino acid substitution” refers to an amino acidsubstitution that does not alter the relative charge or sizecharacteristics of the protein in which the amino acid substitution ismade. Variants can be prepared according to methods for alteringpolypeptide sequence known to one of ordinary skill in the art such asare found in references which compile such methods, e.g. MolecularCloning: A Laboratory Manual, J. Sambrook, et al., eds., Second Edition,Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y., 1989, orCurrent Protocols in Molecular Biology, F. M. Ausubel, et al., eds.,John Wiley & Sons, Inc., New York. Conservative substitutions of aminoacids include substitutions made amongst amino acids within thefollowing groups: (a) M, I, L, V; (b) F, Y, W; (c) K, R, H; (d) A, G;(e) S, T; (f) Q, N; and (g) E, D.

The “percent identity” of two amino acid sequences is determined usingthe algorithm of Karlin and Altschul Proc. Natl. Acad. Sci. USA87:2264-68, 1990, modified as in Karlin and Altschul Proc. Natl. Acad.Sci. USA 90:5873-77, 1993. Such an algorithm is incorporated into theNBLAST and XBLAST programs (version 2.0) of Altschul, et al. J. Mol.Biol. 215:403-10, 1990. BLAST protein searches can be performed with theXBLAST program, score=50, wordlength=3 to obtain amino acid sequenceshomologous to the protein molecules of interest. Where gaps existbetween two sequences, Gapped BLAST can be utilized as described inAltschul et al., Nucleic Acids Res. 25(17):3389-3402, 1997. Whenutilizing BLAST and Gapped BLAST programs, the default parameters of therespective programs (e.g., XBLAST and NBLAST) can be used. Theanti-Galectin-9 antibody may comprise a heavy chain variable regionframework derived from a subclass of germline V_(H) fragment. Suchgermline V_(H) regions are well known in the art. See, e.g., the IMGTdatabase (www.imgt.org) or at www.vbase2.org/vbstat.php. Examplesinclude the IGHV1 subfamily (e.g., IGHV1-2, IGHV1-3, IGHV1-8, IGHV1-18,IGHV1-24, IGHV1-45, IGHV1-46, IGHV1-58, and IGHV1-69), the IGHV2subfamily (e.g., IGHV2-5, IGHV2-26, and IGHV2-70), the IGHV3 subfamily(e.g., IGHV3-7, IGHV3-9, IGHV3-11, IGHV3-13, IGHV3-15, IGHV3-20,IGHV3-21, IGHV3-23, IGHV3-30, IGHV3-33, IGHV3-43, IGHV3-48, IGHV3-49,IGHV3-53, IGHV3-64, IGHV3-66, IGHV3-72, and IGHV3-73, IGHV3-74), theIGHV4 subfamily (e.g., IGHV4-4, IGHV4-28, IGHV4-31, IGHV4-34, IGHV4-39,IGHV4-59, IGHV4-61, and IGHV4-B), the IGHV subfamily (e.g., IGHV5-51, orIGHV6-1), and the IGHV7 subfamily (e.g., IGHV7-4-1).

Alternatively or in addition, the anti-Galectin-9 antibody may comprisea light chain variable region that contains a framework derived from agermline Vκ fragment. Examples include an IGKV1 framework (e.g.,IGKV1-05, IGKV1-12, IGKV1-27, IGKV1-33, or IGKV1-39), an IGKV2 framework(e.g., IGKV2-28), an IGKV3 framework (e.g., IGKV3-11, IGKV3-15, orIGKV3-20), and an IGKV4 framework (e.g., IGKV4-1). In other instances,the anti-Galectin-9 antibody may comprise a light chain variable regionthat contains a framework derived from a germline Vλ, fragment. Examplesinclude an IGλ1 framework (e.g., IGλV1-36, IGλV1-40, IGλV1-44, IGλV1-47,IGλV1-51), an IGλ2 framework (e.g., IGλ, V2-8, IGλV2-11, IGλV2-14,IGλV2-18, IGλV2-23,), an IGλ3 framework (e.g., IGλ, V3-1, IGλV3-10,IGλV3-12, IGλV3-16, IGλV3-19, IGλV3-21, IGλV3-25, IGλV3-27,), an IGλ4framework (e.g., IGλV4-60, IGλV4-69,), an IGλ5 framework (e.g.,IGλV5-39, IGλV5-45,), an IGλ6 framework (e.g., IGλV6-57,), an IGλ7framework (e.g., IGλV7-43, IGλV7-46,), an IGλ8 framework (e.g.,IGλV8-61), an IGλ9 framework (e.g., IGλV9-49), or an IGλ10 framework(e.g., IGλ, V10-54).

In some embodiments, the heavy chain of any of the anti-Galectin-9antibodies as described herein may further comprise a heavy chainconstant region (CH) or a portion thereof (e.g., CH1, CH2, CH3, or acombination thereof). The heavy chain constant region can be of anysuitable origin, e.g., human, mouse, rat, or rabbit. In one specificexample, the heavy chain constant region is from a human IgG (a gammaheavy chain) of any IgG subfamily as described herein.

In some embodiments, the heavy chain constant region of the antibodiesdescribed herein may comprise a single domain (e.g., CH1, CH2, or CH3)or a combination of any of the single domains, of a constant region(e.g., SEQ ID NO: 419-423). In some embodiments, the light chainconstant region of the antibodies described herein may comprise a singledomain (e.g., CL), of a constant region (e.g., SEQ ID NO: 418).Exemplary light and heavy chain sequences are listed below. The hIgG1LALA sequence includes two mutations, L234A and L235A, which suppressFcgR binding as well as a P329G mutation to abolish complement C1qbinding, thus abolishing all immune effector functions. These mutationsare underlined and bolded in the sequences listed below. The hIgG4 FabArm Exchange Mutant sequence includes a mutation to suppress Fab ArmExchange (S228P), underlined and bolded. The light chain sequence forG9.2-17 is identical among all G9.-2-17 constructs. Similarly, the lightchain sequence for G9.1-8m13 is identical among all G9.1-8m13constructs. Bolded residues are the VH and VL regions. A IL2 signalsequence (MYRMQLLSCIALSLALVTNS; SEQ ID NO: 29) is located N-terminallyof the variable region. It is used in expression vectors, which iscleaved during secretion and thus not in the mature antibody molecule.The mature protein (after secretion) starts with “EVQ” for the heavychain and “DIM” for the light chain.

Exemplary Heavy and Light Chain Sequences

G9.2-17 hIgG1 Heavy Chain (SEQ ID NO: 157)EVQLVESGGGLVQPGGSLRLSCAASGFIVSSSSIHWVRQAPGKGLEWVAYISSSSGYTYYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARYWSYPSWWPYRGMDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK* _Light Chain (SEQ ID NO: 108)DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQSSTDPITEGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC* G9.2-17 hIgG1 LALA Heavy Chain (SEQ ID NO: 210)EVQLVESGGGLVQPGGSLRLSCAASGFTVSSSSIHWVRQAPGKGLEWVAYISSSSGYTYYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARYWSYPSWWPYRGMDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPE AA GGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKAL G APIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK* Light Chain (SEQ ID NO: 108)DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQSSTDPITEGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC* G9.2-17 hIgG4 Heavy Chain  (SEQ ID NO: 263)EVQLVESGGGLVQPGGSLRLSCAASGFTVSSSSIHWVRQAPGKGLEWVAYISSSSGYTYYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARYWSYPSWWPYRGMDYWGQGTLVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPSCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLIVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSPGK* Light Chain (SEQ ID NO: 108)DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQSSTDPITEGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC* G9.2-17 hIgG4 Fab Arm Exchange mut Heavy Chain (SEQ ID NO: 316)EVQLVESGGGLVQPGGSLRLSCAASGFTVSSSSIHWVRQAPGKGLEWVAYISSSSGYTYYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARYWSYPSWWPYRGMDYWGQGTLVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCP P CPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLIVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSPGK* Light Chain (SEQ ID NO: 108)DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQSSTDPITEGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC* G9.1-8m13 hIgG1 Heavy Chain (SEQ ID NO: 136)EVQLVESGGGLVQPGGSLRLSCAASGFTVSSSSIHWVRQAPGKGLEWVAYTYPYSSSSSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARYSTYSSKWVWGMDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLIVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK* Light Chain (SEQ ID NO: 108)DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQSYYDSNPITEGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC* G9.1-8m13 hIgG1 LALA Heavy Chain (SEQ ID NO: 189)EVQLVESGGGLVQPGGSLRLSCAASGFTVSSSSIHWVRQAPGKGLEWVAYTYPYSSSSSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARYSTYSSKWVWGMDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPE AA GGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLIVLHQDWLNGKEYKCKVSNKAL G APIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK* _Light Chain (SEQ ID NO: 108)DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQSYYDSNPITEGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC* G9.1-8m13 hIgG4 Heavy Chain (SEQ ID NO: 242)EVQLVESGGGLVQPGGSLRLSCAASGFTVSSSSIHWVRQAPGKGLEWVAYTYPYSSSSSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARYSTYSSKWVWGMDYWGQGTLVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPSCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSPGK* Light Chain (SEQ ID NO: 108)DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQSYYDSNPITEGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC* G9.1-8m13 hIgG4 Fab Arm Exchange mut Heavy Chain(SEQ ID NO: 295)EVQLVESGGGLVQPGGSLRLSCAASGFTVSSSSIHWVRQAPGKGLEWVAYTYPYSSSSSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARYSTYSSKWVWGMDYWGQGTLVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSPGK* Light Chain (SEQ ID NO: 108)DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQSYYDSNPITEGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC

In some embodiments, the anti-Galectin 9 antibody has a light chaincomprising, consisting essentially of, or consisting of SEQ ID NO: 108.In some embodiments, the anti-Galectin 9 antibody has a heavy chaincomprising, consisting essentially of, or consisting of any one of thesequences selected from the group consisting of SEQ ID NO: 295, 242,189, 157, 210, 263, 316, and 136. In some embodiments, the anti-Galectin9 antibody has a light chain comprising, consisting essentially of, orconsisting of SEQ ID NO: 108 and a heavy chain comprising, consistingessentially of, or consisting of any one of the sequences selected fromthe group consisting of SEQ ID NO: 295, 242, 189, 157, 210, 263, 316,and 136. In some embodiments, the anti-Galectin 9 antibody has a lightchain comprising SEQ ID NO: 108 and a heavy chain comprising any one ofthe sequences selected from the group consisting of SEQ ID NO: 295, 242,189, 157, 210, 263, 316, and 136. In some embodiments, the anti-Galectin9 antibody has a light chain consisting essentially of SEQ ID NO: 108and a heavy chain consisting essentially of any one of the sequencesselected from the group consisting of SEQ ID NO: 295, 242, 189, 157,210, 263, 316, and 136. In some embodiments, the anti-Galectin 9antibody has a light chain consisting of SEQ ID NO: 108 and a heavychain consisting of any one of the sequences selected from the groupconsisting of SEQ ID NO: 295, 242, 189, 157, 210, 263, 316, and 136.

In some embodiments, the constant region is from human IgG4. In oneembodiment, the constant region of the anti-Galectin-9 antibodycomprises a heavy chain IgG4 constant region that has at least 80%(e.g., 85%, 90%, 95%, 96%, 97%, 98%, 99% and any increment therein)sequence identity to SEQ ID NO: 423. In one embodiment, the constantregion of the anti-Galectin-9 antibody comprises a heavy chain IgG4constant region comprising SEQ ID NO: 423. In one embodiment, theconstant region of the anti-Galectin-9 antibody comprises a heavy chainIgG4 constant region consisting of SEQ ID NO: 423. In one embodiment,the constant region of the anti-Galectin-9 antibody comprises a heavychain IgG4 constant region that has at least 80% (e.g., 85%, 90%, 95%,96%, 97%, 98%, 99% and any increment therein) sequence identity to SEQID NO: 421. In one embodiment, the constant region of theanti-Galectin-9 antibody comprises a heavy chain IgG4 constant regioncomprising SEQ ID NO: 421. In one embodiment, the constant region of theanti-Galectin-9 antibody comprises a heavy chain IgG4 constant regionconsisting of SEQ ID NO: 421.

In some embodiments, the constant region of the anti-Galectin-9 antibodycomprises a light chain IgG4 constant region that has at least 80%(e.g., 85%, 90%, 95%, 96%, 97%, 98%, 99% and any increment therein)sequence identity to SEQ ID NO: 418. In one embodiment, the constantregion of the anti-Galectin-9 antibody comprises a light chain IgG4constant region comprising SEQ ID No: 418. In one embodiment, theconstant region of the anti-Galectin-9 antibody comprises a light chainIgG4 constant region consisting of SEQ ID NO: 418. In some embodiments,the constant region is from a human IgG1. In some embodiments, theconstant region of the anti-Galectin-9 antibody comprises a heavy chainIgG1 constant region that has at least 80% (e.g., 85%, 90%, 95%, 96%,97%, 98%, 99% and any increment therein) sequence identity to SEQ ID NO:419. In one embodiment, the constant region of the anti-Galectin-9antibody comprises a heavy chain IgG1 constant region comprising SEQ IDNO: 419. In one embodiment, the constant region of the anti-Galectin-9antibody comprises a heavy chain IgG4 constant region consisting of SEQID NO: 419. In one embodiment, the constant region of theanti-Galectin-9 antibody comprises a light chain IgG1 constant regioncomprising SEQ ID NO: 418. In one embodiment, the constant region of theanti-Galectin-9 antibody comprises a light chain IgG4 constant regionconsisting of SEQ ID NO: 418.

In some embodiments, the anti-Galectin-9 antibody comprises a modifiedconstant region. In some embodiments, the anti-Galectin-9 antibodycomprise a modified constant region that is immunologically inert, e.g.,does not trigger complement mediated lysis, or does not stimulateantibody-dependent cell mediated cytotoxicity (ADCC). ADCC activity canbe assessed using methods disclosed in U.S. Pat. No. 5,500,362. In otherembodiments, the constant region is modified as described in Eur. J.Immunol. (1999) 29:2613-2624; PCT Application No. PCT/GB99/01441; and/orUK Patent Application No. 9809951.8. In some embodiments, the IgG4constant region is a mutant with reduced heavy chain exchange. In someembodiments, the constant region is from a human IgG4 Fab Arm Exchangemutant S228P. In one embodiment, the constant region of theanti-Galectin-9 antibody comprises a heavy chain IgG4 constant regionthat has at least 80% (e.g., 85%, 90%, 95%, 96%, 97%, 98%, 99% and anyincrement therein) sequence identity to SEQ ID NO: 422. In oneembodiment, the constant region of the anti-Galectin-9 antibodycomprises a heavy chain IgG4 constant region comprising SEQ ID NO: 422.In one embodiment, the constant region of the anti-Galectin-9 antibodycomprises a heavy chain IgG4 constant region consisting of SEQ ID NO:422. In some embodiments, the constant region of the anti-Galectin-9antibody comprises a light chain IgG4 constant region that has at least80% (e.g., 85%, 90%, 95%, 96%, 97%, 98%, 99% and any increment therein)sequence identity to SEQ ID NO: 418. In one embodiment, the constantregion of the anti-Galectin-9 antibody comprises a light chain IgG4constant region comprising SEQ ID NO: 418. In one embodiment, theconstant region of the anti-Galectin-9 antibody comprises a light chainIgG4 constant region consisting of SEQ ID NO: 418. In some embodiments,the IgG is a mutant with minimal Fc receptor engagement. In one example,the constant region is from a human IgG1 LALA. In one embodiment, theconstant region of the anti-Galectin-9 antibody comprises a heavy chainIgG1 constant region that has at least 80% (e.g., 85%, 90%, 95%, 96%,97%, 98%, 99% and any increment therein) sequence identity to SEQ ID NO:420. In one embodiment, the constant region of the anti-Galectin-9antibody comprises a heavy chain IgG1 constant region comprising SEQ IDNO: 420. In one embodiment, the constant region of the anti-Galectin-9antibody comprises a heavy chain IgG1 constant region consisting of SEQID NO: 420. In one embodiment, the constant region of theanti-Galectin-9 antibody comprises a light chain IgG1 constant regionthat has at least 80% (e.g., 85%, 90%, 95%, 96%, 97%, 98%, 99% and anyincrement therein) sequence identity to SEQ ID NO: 418. In oneembodiment, the constant region of the anti-Galectin-9 antibodycomprises a light chain IgG1 constant region comprising SEQ ID NO: 418.In one embodiment, the constant region of the anti-Galectin-9 antibodycomprises a light chain IgG4 constant region consisting of SEQ ID NO:418.

In some embodiments, the anti-Galectin-9 antibody has chainscorresponding to SEQ ID NO: 88-98 (anti-Galectin-9 antibodies binding toCRD1) and SEQ ID NO: 99-115 (anti-Galectin-9 antibodies binding to CRD2)for the light chains; The amino acid sequences of exemplary heavy chainscorrespond to SEQ ID NO: 116-140 (hIgG1); 169-193 (hIgG1 LALA); 222-246(hIgG4); 275-299 (hIgG4 exchange mut) (anti-Galectin-9 antibodiesbinding to CRD1) and SEQ ID NO: 141-168 (hIgG1); 194-221 (hIgG1 LALA);247-274 (hIgG4); 300-327 (hIgG4 exchange mut) (anti-Galectin-9antibodies binding to CRD2) for the heavy chains. IgG LALA, IgG4exchange mut are located in the heavy chains; accordingly the lightchains are the same for all IgG1 and IgG4 sequences disclosed herein. Insome embodiments, the amino acid sequences of exemplary anti-Galectinantibody light chains correspond to sequences set forth in SEQ ID NO:88-98 and SEQ ID NO: 99-115.

Clone 9.1-Derived Light Chains

In some embodiments, light chains of anti-Galectin-9 antibodies comprisean amino acid sequence that has at least 80% (e.g., 85%, 90%, 95%, 96%,97%, 98%, 99% and any increment therein) sequence identity to any of thelight chains set forth herein (or their variable regions), (e.g., lightchain sequences set forth in SEQ ID NO: 88-98. In some embodiments,light chains of anti-Galectin-9 antibodies consist of an amino acidsequence that has at least 80% (e.g., 85%, 90%, 95%, 96%, 97%, 98%, 99%and any increment therein) sequence identity to any of the light chainsset forth herein, (e.g., light chain sequences set forth in SEQ ID NO:88-98. In some embodiments, light chains of anti-Galectin-9 antibodiescomprise an amino acid sequence set forth in SEQ ID NO: 88-98. In someembodiments, light chains of anti-Galectin-9 antibodies consist of anamino acid sequence set forth in SEQ ID NO: 88-98.

In some embodiments, the anti-Galectin-9 antibodies or antigen-bindingportion thereof comprise a light chain sequence of SEQ ID NO: 88. Insome embodiments, the anti-Galectin-9 antibodies or antigen-bindingportion thereof comprise a light chain sequence of SEQ ID NO: 89. Insome embodiments, the anti-Galectin-9 antibodies or antigen-bindingportion thereof comprise a light chain sequence of SEQ ID NO: 90. Insome embodiments, the anti-Galectin-9 antibodies or antigen-bindingportion thereof comprise a light chain sequence of SEQ ID NO: 91. Insome embodiments, the anti-Galectin-9 antibodies or antigen-bindingportion thereof comprise a light chain sequence of SEQ ID NO: 92. Insome embodiments, the anti-Galectin-9 antibodies or antigen-bindingportion thereof comprise a light chain sequence of SEQ ID NO: 93. Insome embodiments, the anti-Galectin-9 antibodies or antigen-bindingportion thereof comprise a light chain sequence of SEQ ID NO: 94. Insome embodiments, the anti-Galectin-9 antibodies or antigen-bindingportion thereof comprise a light chain sequence of SEQ ID NO: 95. Insome embodiments, the anti-Galectin-9 antibodies or antigen-bindingportion thereof comprise a light chain sequence of SEQ ID NO: 96. Insome embodiments, the anti-Galectin-9 antibodies or antigen-bindingportion thereof comprise a light chain sequence of SEQ ID NO: 97. Insome embodiments, the anti-Galectin-9 antibodies or antigen-bindingportion thereof comprise a light chain sequence of SEQ ID NO: 98.

In some embodiments, light chains of anti-Galectin-9 antibodies comprisean amino acid sequence that has at least 80% (e.g., 85%, 90%, 95%, 96%,97%, 98%, 99% and any increment therein) sequence identity to the lightchain sequence set forth in SEQ ID NO: 95 (or their variable regions).In some embodiments, light chains of anti-Galectin-9 antibodies consistof an amino acid sequence that has at least 80% (e.g., 85%, 90%, 95%,96%, 97%, 98%, 99% and any increment therein) sequence identity to thelight chain sequence set forth in SEQ ID NO: 95. In some embodiments,light chains of anti-Galectin-9 antibodies comprise an amino acidsequence set forth in SEQ ID NO: 95 (or their variable regions). In someembodiments, light chains of anti-Galectin-9 antibodies consist of theamino acid sequence set forth in SEQ ID NO: 95.

Clone 9.1-Derived Heavy Chains

In some embodiments, the amino acid sequences of exemplary anti-Galectinantibody heavy chains correspond to sequences set forth in SEQ ID NO:116-140 (hIgG1); 169-193 (hIgG1 LALA); 222-246 (hIgG4); 275-299 (hIgG4exchange mut) (anti-Galectin-9 antibodies binding to CRD1). In someembodiments, heavy chains of anti-Galectin-9 antibodies comprise anamino acid sequence that has at least 80% (e.g., 85%, 90%, 95%, 96%,97%, 98%, 99% and any increment therein) sequence identity to any of theheavy chains set forth herein (or their variable regions), e.g.,sequences set forth in SEQ ID NO: 116-140; 169-193; 222-246; 275-299(anti-Galectin-9 antibodies binding to CRD1). In some embodiments, heavychains of anti-Galectin-9 antibodies consist of an amino acid sequencethat has at least 80% (e.g., 85%, 90%, 95%, 96%, 97%, 98%, 99% and anyincrement therein) sequence identity to any of the heavy chains setforth herein, e.g., sequences set forth in SEQ ID NO: 116-140; 169-193;222-246; 275-299 (anti-Galectin-9 antibodies binding to CRD1).

In some embodiments, heavy chains of anti-Galectin-9 antibodies comprisean heavy chain amino acid sequence set forth in SEQ ID NO: 116-140;169-193; 222-246; 275-299 (anti-Galectin-9 antibodies binding to CRD1).In some embodiments, heavy chains of anti-Galectin-9 antibodies consistof an amino acid sequence set forth in SEQ ID NO: 116-140; 169-193;222-246; 275-299 (anti-Galectin-9 antibodies binding to CRD1).

In some embodiments, the constant region is IgG1. In some embodiments,the anti-Galectin-9 antibodies or antigen-binding portion thereofcomprise a heavy chain sequence of SEQ ID NO: 116. In some embodiments,the anti-Galectin-9 antibodies or antigen-binding portion thereofcomprise a heavy chain sequence of SEQ ID NO: 117. In some embodiments,the anti-Galectin-9 antibodies or antigen-binding portion thereofcomprise a heavy chain sequence of SEQ ID NO: 118. In some embodiments,the anti-Galectin-9 antibodies or antigen-binding portion thereofcomprise a heavy chain sequence of SEQ ID NO: 119. In some embodiments,the anti-Galectin-9 antibodies or antigen-binding portion thereofcomprise a heavy chain sequence of SEQ ID NO: 120. In some embodiments,the anti-Galectin-9 antibodies or antigen-binding portion thereofcomprise a heavy chain sequence of SEQ ID NO: 121. In some embodiments,the anti-Galectin-9 antibodies or antigen-binding portion thereofcomprise a heavy chain sequence of SEQ ID NO: 122. In some embodiments,the anti-Galectin-9 antibodies or antigen-binding portion thereofcomprise a heavy chain sequence of SEQ ID NO: 123. In some embodiments,the anti-Galectin-9 antibodies or antigen-binding portion thereofcomprise a heavy chain sequence of SEQ ID NO: 124. In some embodiments,the anti-Galectin-9 antibodies or antigen-binding portion thereofcomprise a heavy chain sequence of SEQ ID NO: 125. In some embodiments,the anti-Galectin-9 antibodies or antigen-binding portion thereofcomprise a heavy chain sequence of SEQ ID NO: 126. In some embodiments,the anti-Galectin-9 antibodies or antigen-binding portion thereofcomprise a heavy chain sequence of SEQ ID NO: 127. In some embodiments,the anti-Galectin-9 antibodies or antigen-binding portion thereofcomprise a heavy chain sequence of SEQ ID NO: 128. In some embodiments,the anti-Galectin-9 antibodies or antigen-binding portion thereofcomprise a heavy chain sequence of SEQ ID NO: 129. In some embodiments,the anti-Galectin-9 antibodies or antigen-binding portion thereofcomprise a heavy chain sequence of SEQ ID NO: 130. In some embodiments,the anti-Galectin-9 antibodies or antigen-binding portion thereofcomprise a heavy chain sequence of SEQ ID NO: 131. In some embodiments,the anti-Galectin-9 antibodies or antigen-binding portion thereofcomprise a heavy chain sequence of SEQ ID NO: 132. In some embodiments,the anti-Galectin-9 antibodies or antigen-binding portion thereofcomprise a heavy chain sequence of SEQ ID NO: 133. In some embodiments,the anti-Galectin-9 antibodies or antigen-binding portion thereofcomprise a heavy chain sequence of SEQ ID NO: 134. In some embodiments,the anti-Galectin-9 antibodies or antigen-binding portion thereofcomprise a heavy chain sequence of SEQ ID NO: 135. In some embodiments,the anti-Galectin-9 antibodies or antigen-binding portion thereofcomprise a heavy chain sequence of SEQ ID NO: 136. In some embodiments,the anti-Galectin-9 antibodies or antigen-binding portion thereofcomprise a heavy chain sequence of SEQ ID NO: 137. In some embodiments,the anti-Galectin-9 antibodies or antigen-binding portion thereofcomprise a heavy chain sequence of SEQ ID NO: 138. In some embodiments,the anti-Galectin-9 antibodies or antigen-binding portion thereofcomprise a heavy chain sequence of SEQ ID NO: 139. In some embodiments,the anti-Galectin-9 antibodies or antigen-binding portion thereofcomprise a heavy chain sequence of SEQ ID NO: 140.

In some embodiments, the constant region is IgG1 LALA. In someembodiments, the anti-Galectin-9 antibodies or antigen-binding portionthereof comprise a heavy chain sequence of SEQ ID NO: 169. In someembodiments, the anti-Galectin-9 antibodies or antigen-binding portionthereof comprise a heavy chain sequence of SEQ ID NO: 170. In someembodiments, the anti-Galectin-9 antibodies or antigen-binding portionthereof comprise a heavy chain sequence of SEQ ID NO: 171. In someembodiments, the anti-Galectin-9 antibodies or antigen-binding portionthereof comprise a heavy chain sequence of SEQ ID NO: 172. In someembodiments, the anti-Galectin-9 antibodies or antigen-binding portionthereof comprise a heavy chain sequence of SEQ ID NO: 173. In someembodiments, the anti-Galectin-9 antibodies or antigen-binding portionthereof comprise a heavy chain sequence of SEQ ID NO: 174. In someembodiments, the anti-Galectin-9 antibodies or antigen-binding portionthereof comprise a heavy chain sequence of SEQ ID NO: 175. In someembodiments, the anti-Galectin-9 antibodies or antigen-binding portionthereof comprise a heavy chain sequence of SEQ ID NO: 176. In someembodiments, the anti-Galectin-9 antibodies or antigen-binding portionthereof comprise a heavy chain sequence of SEQ ID NO: 177. In someembodiments, the anti-Galectin-9 antibodies or antigen-binding portionthereof comprise a heavy chain sequence of SEQ ID NO: 178. In someembodiments, the anti-Galectin-9 antibodies or antigen-binding portionthereof comprise a heavy chain sequence of SEQ ID NO: 179. In someembodiments, the anti-Galectin-9 antibodies or antigen-binding portionthereof comprise a heavy chain sequence of SEQ ID NO: 180. In someembodiments, the anti-Galectin-9 antibodies or antigen-binding portionthereof comprise a heavy chain sequence of SEQ ID NO: 181. In someembodiments, the anti-Galectin-9 antibodies or antigen-binding portionthereof comprise a heavy chain sequence of SEQ ID NO: 182. In someembodiments, the anti-Galectin-9 antibodies or antigen-binding portionthereof comprise a heavy chain sequence of SEQ ID NO: 183. In someembodiments, the anti-Galectin-9 antibodies or antigen-binding portionthereof comprise a heavy chain sequence of SEQ ID NO: 184. In someembodiments, the anti-Galectin-9 antibodies or antigen-binding portionthereof comprise a heavy chain sequence of SEQ ID NO: 185. In someembodiments, the anti-Galectin-9 antibodies or antigen-binding portionthereof comprise a heavy chain sequence of SEQ ID NO: 186. In someembodiments, the anti-Galectin-9 antibodies or antigen-binding portionthereof comprise a heavy chain sequence of SEQ ID NO: 187. In someembodiments, the anti-Galectin-9 antibodies or antigen-binding portionthereof comprise a heavy chain sequence of SEQ ID NO: 188. In someembodiments, the anti-Galectin-9 antibodies or antigen-binding portionthereof comprise a heavy chain sequence of SEQ ID NO: 189. In someembodiments, the anti-Galectin-9 antibodies or antigen-binding portionthereof comprise a heavy chain sequence of SEQ ID NO: 190. In someembodiments, the anti-Galectin-9 antibodies or antigen-binding portionthereof comprise a heavy chain sequence of SEQ ID NO: 191. In someembodiments, the anti-Galectin-9 antibodies or antigen-binding portionthereof comprise a heavy chain sequence of SEQ ID NO: 192. In someembodiments, the anti-Galectin-9 antibodies or antigen-binding portionthereof comprise a heavy chain sequence of SEQ ID NO: 193.

In some embodiments, the constant region is IgG4. In some embodiments,the anti-Galectin-9 antibodies or antigen-binding portion thereofcomprise a heavy chain sequence of SEQ ID NO: 222. In some embodiments,the anti-Galectin-9 antibodies or antigen-binding portion thereofcomprise a heavy chain sequence of SEQ ID NO: 223. In some embodiments,the anti-Galectin-9 antibodies or antigen-binding portion thereofcomprise a heavy chain sequence of SEQ ID NO: 224. In some embodiments,the anti-Galectin-9 antibodies or antigen-binding portion thereofcomprise a heavy chain sequence of SEQ ID NO: 225. In some embodiments,the anti-Galectin-9 antibodies or antigen-binding portion thereofcomprise a heavy chain sequence of SEQ ID NO: 226. In some embodiments,the anti-Galectin-9 antibodies or antigen-binding portion thereofcomprise a heavy chain sequence of SEQ ID NO: 227. In some embodiments,the anti-Galectin-9 antibodies or antigen-binding portion thereofcomprise a heavy chain sequence of SEQ ID NO: 228. In some embodiments,the anti-Galectin-9 antibodies or antigen-binding portion thereofcomprise a heavy chain sequence of SEQ ID NO: 229. In some embodiments,the anti-Galectin-9 antibodies or antigen-binding portion thereofcomprise a heavy chain sequence of SEQ ID NO: 230. In some embodiments,the anti-Galectin-9 antibodies or antigen-binding portion thereofcomprise a heavy chain sequence of SEQ ID NO: 231. In some embodiments,the anti-Galectin-9 antibodies or antigen-binding portion thereofcomprise a heavy chain sequence of SEQ ID NO: 232. In some embodiments,the anti-Galectin-9 antibodies or antigen-binding portion thereofcomprise a heavy chain sequence of SEQ ID NO: 233. In some embodiments,the anti-Galectin-9 antibodies or antigen-binding portion thereofcomprise a heavy chain sequence of SEQ ID NO: 234. In some embodiments,the anti-Galectin-9 antibodies or antigen-binding portion thereofcomprise a heavy chain sequence of SEQ ID NO: 235. In some embodiments,the anti-Galectin-9 antibodies or antigen-binding portion thereofcomprise a heavy chain sequence of SEQ ID NO: 236. In some embodiments,the anti-Galectin-9 antibodies or antigen-binding portion thereofcomprise a heavy chain sequence of SEQ ID NO: 237. In some embodiments,the anti-Galectin-9 antibodies or antigen-binding portion thereofcomprise a heavy chain sequence of SEQ ID NO: 238. In some embodiments,the anti-Galectin-9 antibodies or antigen-binding portion thereofcomprise a heavy chain sequence of SEQ ID NO: 29. In some embodiments,the anti-Galectin-9 antibodies or antigen-binding portion thereofcomprise a heavy chain sequence of SEQ ID NO: 240. In some embodiments,the anti-Galectin-9 antibodies or antigen-binding portion thereofcomprise a heavy chain sequence of SEQ ID NO: 241. In some embodiments,the anti-Galectin-9 antibodies or antigen-binding portion thereofcomprise a heavy chain sequence of SEQ ID NO: 242. In some embodiments,the anti-Galectin-9 antibodies or antigen-binding portion thereofcomprise a heavy chain sequence of SEQ ID NO: 243. In some embodiments,the anti-Galectin-9 antibodies or antigen-binding portion thereofcomprise a heavy chain sequence of SEQ ID NO: 244. In some embodiments,the anti-Galectin-9 antibodies or antigen-binding portion thereofcomprise a heavy chain sequence of SEQ ID NO: 245. In some embodiments,the anti-Galectin-9 antibodies or antigen-binding portion thereofcomprise a heavy chain sequence of SEQ ID NO: 246.

In some embodiments, the constant region is IgG4mut. In someembodiments, the anti-Galectin-9 antibodies or antigen-binding portionthereof comprise a heavy chain sequence of SEQ ID NO: 275. In someembodiments, the anti-Galectin-9 antibodies or antigen-binding portionthereof comprise a heavy chain sequence of SEQ ID NO: 276. In someembodiments, the anti-Galectin-9 antibodies or antigen-binding portionthereof comprise a heavy chain sequence of SEQ ID NO: 277. In someembodiments, the anti-Galectin-9 antibodies or antigen-binding portionthereof comprise a heavy chain sequence of SEQ ID NO: 278. In someembodiments, the anti-Galectin-9 antibodies or antigen-binding portionthereof comprise a heavy chain sequence of SEQ ID NO: 279. In someembodiments, the anti-Galectin-9 antibodies or antigen-binding portionthereof comprise a heavy chain sequence of SEQ ID NO: 280. In someembodiments, the anti-Galectin-9 antibodies or antigen-binding portionthereof comprise a heavy chain sequence of SEQ ID NO: 281. In someembodiments, the anti-Galectin-9 antibodies or antigen-binding portionthereof comprise a heavy chain sequence of SEQ ID NO: 282. In someembodiments, the anti-Galectin-9 antibodies or antigen-binding portionthereof comprise a heavy chain sequence of SEQ ID NO: 283. In someembodiments, the anti-Galectin-9 antibodies or antigen-binding portionthereof comprise a heavy chain sequence of SEQ ID NO: 284. In someembodiments, the anti-Galectin-9 antibodies or antigen-binding portionthereof comprise a heavy chain sequence of SEQ ID NO: 285. In someembodiments, the anti-Galectin-9 antibodies or antigen-binding portionthereof comprise a heavy chain sequence of SEQ ID NO: 286. In someembodiments, the anti-Galectin-9 antibodies or antigen-binding portionthereof comprise a heavy chain sequence of SEQ ID NO: 287. In someembodiments, the anti-Galectin-9 antibodies or antigen-binding portionthereof comprise a heavy chain sequence of SEQ ID NO: 288. In someembodiments, the anti-Galectin-9 antibodies or antigen-binding portionthereof comprise a heavy chain sequence of SEQ ID NO: 289. In someembodiments, the anti-Galectin-9 antibodies or antigen-binding portionthereof comprise a heavy chain sequence of SEQ ID NO: 290. In someembodiments, the anti-Galectin-9 antibodies or antigen-binding portionthereof comprise a heavy chain sequence of SEQ ID NO: 291. In someembodiments, the anti-Galectin-9 antibodies or antigen-binding portionthereof comprise a heavy chain sequence of SEQ ID NO: 292. In someembodiments, the anti-Galectin-9 antibodies or antigen-binding portionthereof comprise a heavy chain sequence of SEQ ID NO: 293 In someembodiments, the anti-Galectin-9 antibodies or antigen-binding portionthereof comprise a heavy chain sequence of SEQ ID NO: 294. In someembodiments, the anti-Galectin-9 antibodies or antigen-binding portionthereof comprise a heavy chain sequence of SEQ ID NO: 295. In someembodiments, the anti-Galectin-9 antibodies or antigen-binding portionthereof comprise a heavy chain sequence of SEQ ID NO: 296. In someembodiments, the anti-Galectin-9 antibodies or antigen-binding portionthereof comprise a heavy chain sequence of SEQ ID NO: 297. In someembodiments, the anti-Galectin-9 antibodies or antigen-binding portionthereof comprise a heavy chain sequence of SEQ ID NO: 298. In someembodiments, the anti-Galectin-9 antibodies or antigen-binding portionthereof comprise a heavy chain sequence of SEQ ID NO: 299.

In some embodiments, heavy chains of anti-Galectin-9 antibodies comprisean amino acid sequence that has at least 80% (e.g., 85%, 90%, 95%, 96%,97%, 98%, 99% and any increment therein) sequence identity to the heavychain sequence set forth in SEQ ID NO: 136 (or its variable region). Insome embodiments, heavy chains of anti-Galectin-9 antibodies consist ofan amino acid sequence that has at least 80% (e.g., 85%, 90%, 95%, 96%,97%, 98%, 99% and any increment therein) sequence identity to the heavychain sequence set forth in SEQ ID NO: 136. In some embodiments, heavychains of anti-Galectin-9 antibodies comprise an amino acid sequence setforth in SEQ ID NO: 136 (or its variable region). In some embodiments,heavy chains of anti-Galectin-9 antibodies consist of an amino acidsequence set forth in SEQ ID NO: 136.

In some embodiments, heavy chains of anti-Galectin-9 antibodies comprisean amino acid sequence that has at least 80% (e.g., 85%, 90%, 95%, 96%,97%, 98%, 99% and any increment therein) sequence identity to the heavychain sequence set forth in SEQ ID NO: 189 (or its variable region). Insome embodiments, heavy chains of anti-Galectin-9 antibodies consist ofan amino acid sequence that has at least 80% (e.g., 85%, 90%, 95%, 96%,97%, 98%, 99% and any increment therein) sequence identity to the heavychain sequence set forth in SEQ ID NO: 189. In some embodiments, heavychains of anti-Galectin-9 antibodies comprise an amino acid sequence setforth in SEQ ID NO: 189 (or its variable region). In some embodiments,heavy chains of anti-Galectin-9 antibodies consist of an amino acidsequence set forth in SEQ ID NO: 189.

In some embodiments, heavy chains of anti-Galectin-9 antibodies comprisean amino acid sequence that has at least 80% (e.g., 85%, 90%, 95%, 96%,97%, 98%, 99% and any increment therein) sequence identity to the heavychain sequence set forth in SEQ ID NO: 242 (or its variable region). Insome embodiments, heavy chains of anti-Galectin-9 antibodies consist ofan amino acid sequence that has at least 80% (e.g., 85%, 90%, 95%, 96%,97%, 98%, 99% and any increment therein) sequence identity to the heavychain sequence set forth in SEQ ID NO: 242. In some embodiments, heavychains of anti-Galectin-9 antibodies comprise an amino acid sequence setforth in SEQ ID NO: 242 (or its variable region). In some embodiments,heavy chains of anti-Galectin-9 antibodies consist of an amino acidsequence set forth in SEQ ID NO: 242.

In some embodiments, heavy chains of anti-Galectin-9 antibodies comprisean amino acid sequence that has at least 80% (e.g., 85%, 90%, 95%, 96%,97%, 98%, 99% and any increment therein) sequence identity to the heavychain sequence set forth in SEQ ID NO: 295 (or its variable region). Insome embodiments, heavy chains of anti-Galectin-9 antibodies consist ofan amino acid sequence that has at least 80% (e.g., 85%, 90%, 95%, 96%,97%, 98%, 99% and any increment therein) sequence identity to the heavychain sequence set forth in SEQ ID NO: 295. In some embodiments, heavychains of anti-Galectin-9 antibodies comprise an amino acid sequence setforth in SEQ ID NO: 295. In some embodiments, heavy chains ofanti-Galectin-9 antibodies consist of an amino acid sequence set forthin SEQ ID NO: 295.

Clone 9.1 Derived Heavy and Light Chains

A VH domain can comprise the amino acid sequence of any VH domaindescribed herein fused to a human IgG, e.g., an IgG1, constant region,such as human IgG1 constant domain amino acid sequence, hIgG LALA,hIgG4, or IgG4mut.

In some embodiments, the amino acid sequences of exemplary anti-Galectinantibody light chains correspond to SEQ ID NO: 88-98, or the amino acidsequences of the exemplary anti-Galectin antibody heavy chainscorrespond to SEQ ID NO: 116-140; 169-193; 222-246; 275-299.

In some embodiments, the anti-Galectin-9 antibodies or antigen-bindingportion thereof comprise a light chain having the sequence of SEQ ID NO:88 and a heavy chain having a sequence selected from of SEQ ID NO: 116,169, 222, or 275.

In some embodiments, the anti-Galectin-9 antibodies or antigen-bindingportion thereof comprise a light chain having the sequence of SEQ ID NO:89 and a heavy chain having a sequence selected from of SEQ ID NO: 117,170, 223, or 276.

In some embodiments, the anti-Galectin-9 antibodies or antigen-bindingportion thereof comprise a light chain having the sequence of SEQ ID NO:90 and a heavy chain having a sequence selected from of SEQ ID NO: 118,171, 224, or 277.

In some embodiments, the anti-Galectin-9 antibodies or antigen-bindingportion thereof comprise a light chain having the sequence of SEQ ID NO:91 and a heavy chain having a sequence selected from of SEQ ID NO: 119,172, 225, or 278.

In some embodiments, the anti-Galectin-9 antibodies or antigen-bindingportion thereof comprise a light chain having the sequence of SEQ ID NO:92 and a heavy chain having a sequence selected from of SEQ ID NO: 120,173, 226, or 279.

In some embodiments, the anti-Galectin-9 antibodies or antigen-bindingportion thereof comprise a light chain having the sequence of SEQ ID NO:93 and a heavy chain having a sequence selected from of SEQ ID NO: 121,174, 227, or 280.

In some embodiments, the anti-Galectin-9 antibodies or antigen-bindingportion thereof comprise a light chain having the sequence of SEQ ID NO:94 and a heavy chain having a sequence selected from of SEQ ID NO: 122,175, 228, or 281.

In some embodiments, the anti-Galectin-9 antibodies or antigen-bindingportion thereof comprise a light chain having the sequence of SEQ ID NO:95 and a heavy chain having a sequence selected from of SEQ ID NO: 123,176, 229, or 282.

In some embodiments, the anti-Galectin-9 antibodies or antigen-bindingportion thereof comprise a light chain having the sequence of SEQ ID NO:96 and a heavy chain having a sequence selected from of SEQ ID NO: 138,191, 244, or 297.

In some embodiments, the anti-Galectin-9 antibodies or antigen-bindingportion thereof comprise a light chain having the sequence of SEQ ID NO:97 and a heavy chain having a sequence selected from of SEQ ID NO: 139,192, 245, or 298.

In some embodiments, the anti-Galectin-9 antibodies or antigen-bindingportion thereof comprise a light chain having the sequence of SEQ ID NO:98 and a heavy chain having a sequence selected from of SEQ ID NO: 140,193, 246, or 299.

In some embodiments, the anti-Galectin-9 antibodies or antigen-bindingportion thereof comprise a light chain having the sequence of SEQ ID NO:95 and a heavy chain having a sequence selected from of SEQ ID NO: 124,177, 230, or 283.

In some embodiments, the anti-Galectin-9 antibodies or antigen-bindingportion thereof comprise a light chain having the sequence of SEQ ID NO:95 and a heavy chain having a sequence selected from of SEQ ID NO: 125,178, 231, or 284.

In some embodiments, the anti-Galectin-9 antibodies or antigen-bindingportion thereof comprise a light chain having the sequence of SEQ ID NO:95 and a heavy chain having a sequence selected from of SEQ ID NO: 126,179, 232, or 285.

In some embodiments, the anti-Galectin-9 antibodies or antigen-bindingportion thereof comprise a light chain having the sequence of SEQ ID NO:95 and a heavy chain having a sequence selected from of SEQ ID NO: 127,180, 233, or 286,

In some embodiments, the anti-Galectin-9 antibodies or antigen-bindingportion thereof comprise a light chain having the sequence of SEQ ID NO:95 and a heavy chain having a sequence selected from of SEQ ID NO: 128,181, 234, or 287.

In some embodiments, the anti-Galectin-9 antibodies or antigen-bindingportion thereof comprise a light chain having the sequence of SEQ ID NO:95 and a heavy chain having a sequence selected from of SEQ ID NO: 129,182, 235, or 288.

In some embodiments, the anti-Galectin-9 antibodies or antigen-bindingportion thereof comprise a light chain having the sequence of SEQ ID NO:95 and a heavy chain having a sequence selected from of SEQ ID NO: 130,183, 236, or 289.

In some embodiments, the anti-Galectin-9 antibodies or antigen-bindingportion thereof comprise a light chain having the sequence of SEQ ID NO:95 and a heavy chain having a sequence selected from of SEQ ID NO: 131,184, 237, or 290.

In some embodiments, the anti-Galectin-9 antibodies or antigen-bindingportion thereof comprise a light chain having the sequence of SEQ ID NO:95 and a heavy chain having a sequence selected from of SEQ ID NO: 132,185, 238, or 291.

In some embodiments, the anti-Galectin-9 antibodies or antigen-bindingportion thereof comprise a light chain having the sequence of SEQ ID NO:95 and a heavy chain having a sequence selected from of SEQ ID NO: 133,186, 239, or 292.

In some embodiments, the anti-Galectin-9 antibodies or antigen-bindingportion thereof comprise a light chain having the sequence of SEQ ID NO:95 and a heavy chain having a sequence selected from of SEQ ID NO: 137,187, 240, or 293.

In some embodiments, the anti-Galectin-9 antibodies or antigen-bindingportion thereof comprise a light chain having the sequence of SEQ ID NO:95 and a heavy chain having a sequence selected from of SEQ ID NO: 138,188, 241, or 294.

In some embodiments, the anti-Galectin-9 antibodies or antigen-bindingportion thereof comprise a light chain having the sequence of SEQ ID NO:95 and a heavy chain having a sequence selected from of SEQ ID NO: 139,189, 242, or 295.

In some embodiments, the anti-Galectin-9 antibodies or antigen-bindingportion thereof comprise a light chain having the sequence of SEQ ID NO:95 and a heavy chain having a sequence selected from of SEQ ID NO: 140,190, 243, or 296.

In some embodiments, the anti-Galectin-9 antibodies or antigen-bindingportion thereof comprise a light chain sequence of SEQ ID NO: 95 andcomprise a heavy chain sequence of SEQ ID NO: 136. In some embodiments,light chains of anti-Galectin-9 antibodies comprise an amino acidsequence that has at least 80% (e.g., 85%, 90%, 95%, 96%, 97%, 98%, 99%and any increment therein) sequence identity to the light chain sequenceset forth in SEQ ID NO: 95 (or their variable regions), and heavy chainsof anti-Galectin-9 antibodies comprise an amino acid sequence that hasat least 80% (e.g., 85%, 90%, 95%, 96%, 97%, 98%, 99% and any incrementtherein) sequence identity to the heavy chain sequence set forth in SEQID NO: 136 (or its variable region). In some embodiments, light chainsof anti-Galectin-9 antibodies consist of an amino acid sequence that hasat least 80% (e.g., 85%, 90%, 95%, 96%, 97%, 98%, 99% and any incrementtherein) sequence identity to the light chain sequence set forth in SEQID NO: 95 and heavy chains of anti-Galectin-9 antibodies consist of anamino acid sequence that has at least 80% (e.g., 85%, 90%, 95%, 96%,97%, 98%, 99% and any increment therein) sequence identity to the heavychain sequence set forth in SEQ ID NO: 136. In some embodiments, lightchains of anti-Galectin-9 antibodies comprise an amino acid sequence setforth in SEQ ID NO: 95 (or their variable regions) heavy chains ofanti-Galectin-9 antibodies comprise an amino acid sequence set forth inSEQ ID NO: 136 (or its variable region). In some embodiments, lightchains of anti-Galectin-9 antibodies consist of the amino acid sequenceset forth in SEQ ID NO: 95 and heavy chains of anti-Galectin-9antibodies consist of an amino acid sequence set forth in SEQ ID NO:136.

In some embodiments, the anti-Galectin-9 antibodies or antigen-bindingportion thereof comprise a light chain sequence of SEQ ID NO: 95 andcomprise a heavy chain sequence of SEQ ID NO: 189. In some embodiments,light chains of anti-Galectin-9 antibodies comprise an amino acidsequence that has at least 80% (e.g., 85%, 90%, 95%, 96%, 97%, 98%, 99%and any increment therein) sequence identity to the light chain sequenceset forth in SEQ ID NO: 95 (or their variable regions) and heavy chainsof anti-Galectin-9 antibodies comprise an amino acid sequence that hasat least 80% (e.g., 85%, 90%, 95%, 96%, 97%, 98%, 99% and any incrementtherein) sequence identity to the heavy chain sequence set forth in SEQID NO: 189 (or its variable region) and heavy chains of anti-Galectin-9antibodies consist of an amino acid sequence that has at least 80%(e.g., 85%, 90%, 95%, 96%, 97%, 98%, 99% and any increment therein)sequence identity to the heavy chain sequence set forth in SEQ ID NO:189. In some embodiments, light chains of anti-Galectin-9 antibodiesconsist of an amino acid sequence that has at least 80% (e.g., 85%, 90%,95%, 96%, 97%, 98%, 99% and any increment therein) sequence identity tothe light chain sequence set forth in SEQ ID NO: 95. In someembodiments, light chains of anti-Galectin-9 antibodies comprise anamino acid sequence set forth in SEQ ID NO: 95 (or their variableregions) and heavy chains of anti-Galectin-9 antibodies comprise anamino acid sequence set forth in SEQ ID NO: 189 (or its variableregion). In some embodiments, light chains of anti-Galectin-9 antibodiesconsist of the amino acid sequence set forth in SEQ ID NO: 95 and heavychains of anti-Galectin-9 antibodies comprise an amino acid sequence setforth in SEQ ID NO: 189.

In some embodiments, the anti-Galectin-9 antibodies or antigen-bindingportion thereof comprise a light chain sequence of SEQ ID NO: 95 andcomprise a heavy chain sequence of SEQ ID NO: 242. In some embodiments,light chains of anti-Galectin-9 antibodies comprise an amino acidsequence that has at least 80% (e.g., 85%, 90%, 95%, 96%, 97%, 98%, 99%and any increment therein) sequence identity to the light chain sequenceset forth in SEQ ID NO: 95 (or their variable regions) and heavy chainsof anti-Galectin-9 antibodies comprise an amino acid sequence that hasat least 80% (e.g., 85%, 90%, 95%, 96%, 97%, 98%, 99% and any incrementtherein) sequence identity to the heavy chain sequence set forth in SEQID NO: 242 (or its variable region). In some embodiments, light chainsof anti-Galectin-9 antibodies consist of an amino acid sequence that hasat least 80% (e.g., 85%, 90%, 95%, 96%, 97%, 98%, 99% and any incrementtherein) sequence identity to the light chain sequence set forth in SEQID NO: 95 and heavy chains of anti-Galectin-9 antibodies consist of anamino acid sequence that has at least 80% (e.g., 85%, 90%, 95%, 96%,97%, 98%, 99% and any increment therein) sequence identity to the heavychain sequence set forth in SEQ ID NO: 242. In some embodiments, lightchains of anti-Galectin-9 antibodies comprise an amino acid sequence setforth in SEQ ID NO: 95 (or their variable regions) and heavy chains ofanti-Galectin-9 antibodies comprise an amino acid sequence set forth inSEQ ID NO: 242 (or its variable region). In some embodiments, lightchains of anti-Galectin-9 antibodies consist of the amino acid sequenceset forth in SEQ ID NO: 95 and heavy chains of anti-Galectin-9antibodies consist of an amino acid sequence set forth in SEQ ID NO:242.

In some embodiments, the anti-Galectin-9 antibodies or antigen-bindingportion thereof comprise a light chain sequence of SEQ ID NO: 95 andcomprise a heavy chain sequence of SEQ ID NO: 295. In some embodiments,light chains of anti-Galectin-9 antibodies comprise an amino acidsequence that has at least 80% (e.g., 85%, 90%, 95%, 96%, 97%, 98%, 99%and any increment therein) sequence identity to the light chain sequenceset forth in SEQ ID NO: 95 (or their variable regions) and heavy chainsof anti-Galectin-9 antibodies comprise an amino acid sequence that hasat least 80% (e.g., 85%, 90%, 95%, 96%, 97%, 98%, 99% and any incrementtherein) sequence identity to the heavy chain sequence set forth in SEQID NO: 295 (or its variable region). In some embodiments, light chainsof anti-Galectin-9 antibodies consist of an amino acid sequence that hasat least 80% (e.g., 85%, 90%, 95%, 96%, 97%, 98%, 99% and any incrementtherein) sequence identity to the light chain sequence set forth in SEQID NO: 95 and heavy chains of anti-Galectin-9 antibodies consist of anamino acid sequence that has at least 80% (e.g., 85%, 90%, 95%, 96%,97%, 98%, 99% and any increment therein) sequence identity to the heavychain sequence set forth in SEQ ID NO: 295. In some embodiments, lightchains of anti-Galectin-9 antibodies comprise an amino acid sequence setforth in SEQ ID NO: 95 (or their variable regions) and heavy chains ofanti-Galectin-9 antibodies comprise an amino acid sequence set forth inSEQ ID NO: 295. In some embodiments, light chains of anti-Galectin-9antibodies consist of the amino acid sequence set forth in SEQ ID NO: 95and heavy chains of anti-Galectin-9 antibodies consist of an amino acidsequence set forth in SEQ ID NO: 295.

Clone 9.2-Derived Light Chains

In some embodiments, light chains of anti-Galectin-9 antibodies comprisean amino acid sequence that has at least 80% (e.g., 85%, 90%, 95%, 96%,97%, 98%, 99% and any increment therein) sequence identity to any of thelight chains set forth herein (or their variable regions), (e.g., lightchain sequences set forth in SEQ ID NO: 99-115). In some embodiments,light chains of anti-Galectin-9 antibodies consist of an amino acidsequence that has at least 80% (e.g., 85%, 90%, 95%, 96%, 97%, 98%, 99%and any increment therein) sequence identity to any of the light chainsset forth herein, (e.g., light chain sequences set forth in SEQ ID NO:99-115).

In some embodiments, light chains of anti-Galectin-9 antibodies comprisean amino acid sequence set forth in SEQ ID NO: 99-115. In someembodiments, light chains of anti-Galectin-9 antibodies consist of asequence set forth in SEQ ID NO: 99-115.

In some embodiments, the anti-Galectin-9 antibodies or antigen-bindingportion thereof comprise a light chain sequence of SEQ ID NO: 99. Insome embodiments, the anti-Galectin-9 antibodies or antigen-bindingportion thereof comprise a light chain sequence of SEQ ID NO: 100. Insome embodiments, the anti-Galectin-9 antibodies or antigen-bindingportion thereof comprise a light chain sequence of SEQ ID NO: 101. Insome embodiments, the anti-Galectin-9 antibodies or antigen-bindingportion thereof comprise a light chain sequence of SEQ ID NO: 102. Insome embodiments, the anti-Galectin-9 antibodies or antigen-bindingportion thereof comprise a light chain sequence of SEQ ID NO: 103. Insome embodiments, the anti-Galectin-9 antibodies or antigen-bindingportion thereof comprise a light chain sequence of SEQ ID NO: 104. Insome embodiments, the anti-Galectin-9 antibodies or antigen-bindingportion thereof comprise a light chain sequence of SEQ ID NO: 105. Insome embodiments, the anti-Galectin-9 antibodies or antigen-bindingportion thereof comprise a light chain sequence of SEQ ID NO: 106. Insome embodiments, the anti-Galectin-9 antibodies or antigen-bindingportion thereof comprise a light chain sequence of SEQ ID NO: 107. Insome embodiments, the anti-Galectin-9 antibodies or antigen-bindingportion thereof comprise a light chain sequence of SEQ ID NO: 108. Insome embodiments, the anti-Galectin-9 antibodies or antigen-bindingportion thereof comprise a light chain sequence of SEQ ID NO: 109. Insome embodiments, the anti-Galectin-9 antibodies or antigen-bindingportion thereof comprise a light chain sequence of SEQ ID NO: 110. Insome embodiments, the anti-Galectin-9 antibodies or antigen-bindingportion thereof comprise a light chain sequence of SEQ ID NO: 111. Insome embodiments, the anti-Galectin-9 antibodies or antigen-bindingportion thereof comprise a light chain sequence of SEQ ID NO: 112. Insome embodiments, the anti-Galectin-9 antibodies or antigen-bindingportion thereof comprise a light chain sequence of SEQ ID NO: 113. Insome embodiments, the anti-Galectin-9 antibodies or antigen-bindingportion thereof comprise a light chain sequence of SEQ ID NO: 114. Insome embodiments, the anti-Galectin-9 antibodies or antigen-bindingportion thereof comprise a light chain sequence of SEQ ID NO: 115. Insome embodiments, light chains of anti-Galectin-9 antibodies comprise anamino acid sequence that has at least 80% (e.g., 85%, 90%, 95%, 96%,97%, 98%, 99% and any increment therein) sequence identity to the lightchain sequence set forth in SEQ ID NO: 108 (or their variable regions).In some embodiments, light chains of anti-Galectin-9 antibodies consistof an amino acid sequence that has at least 80% (e.g., 85%, 90%, 95%,96%, 97%, 98%, 99% and any increment therein) sequence identity to thelight chain sequence set forth in SEQ ID NO: 108.

In some embodiments, light chains of anti-Galectin-9 antibodies comprisean amino acid sequence set forth in SEQ ID NO: 108 (or their variableregions). In some embodiments, light chains of anti-Galectin-9antibodies consist set forth in SEQ ID NO: 108.

Clone 9.2-Derived Heavy Chains

In some embodiments, the amino acid sequences of exemplary anti-Galectinantibody heavy chains correspond to sequences set forth in SEQ ID NO:141-168 (hIgG1); 194-221 (hIgG1 LALA); 247-274 (hIgG4); 300-327 (hIgG4exchange mut) (anti-Galectin-9 antibodies binding to CRD2) for the heavychains.

In some embodiments, heavy chains of anti-Galectin-9 antibodies comprisean amino acid sequence that has at least 80% (e.g., 85%, 90%, 95%, 96%,97%, 98%, 99% and any increment therein) sequence identity to any of theheavy chains set forth herein (or their variable regions), e.g.,sequences set forth in SEQ ID NO: 141-168; 194-220; 247-274; 300-327(anti-Galectin-9 antibodies binding to CRD2). In some embodiments, heavychains of anti-Galectin-9 antibodies consist of an amino acid sequencethat has at least 80% (e.g., 85%, 90%, 95%, 96%, 97%, 98%, 99% and anyincrement therein) sequence identity to any of the heavy chains setforth herein, e.g., sequences set forth in SEQ ID NO: 141-168; 194-220;247-274; 300-327 (anti-Galectin-9 antibodies binding to CRD2).

In some embodiments, heavy chains of anti-Galectin-9 antibodies comprisean heavy chain amino acid sequence set forth in SEQ ID NO: 141-168;194-220; 247-274; 300-327 (anti-Galectin-9 antibodies binding to CRD2).In some embodiments, heavy chains of anti-Galectin-9 antibodies consistof an amino acid sequence set forth in SEQ ID NO: 141-168; 194-220;247-274; 300-327 (anti-Galectin-9 antibodies binding to CRD2).

In some embodiments, the constant region is IgG1. In some embodiments,the anti-Galectin-9 antibodies or antigen-binding portion thereofcomprise a heavy chain sequence of SEQ ID NO: 141. In some embodiments,the anti-Galectin-9 antibodies or antigen-binding portion thereofcomprise a heavy chain sequence of SEQ ID NO: 142. In some embodiments,the anti-Galectin-9 antibodies or antigen-binding portion thereofcomprise a heavy chain sequence of SEQ ID NO: 143. In some embodiments,the anti-Galectin-9 antibodies or antigen-binding portion thereofcomprise a heavy chain sequence of SEQ ID NO: 144. In some embodiments,the anti-Galectin-9 antibodies or antigen-binding portion thereofcomprise a heavy chain sequence of SEQ ID NO: 145. In some embodiments,the anti-Galectin-9 antibodies or antigen-binding portion thereofcomprise a heavy chain sequence of SEQ ID NO: 146. In some embodiments,the anti-Galectin-9 antibodies or antigen-binding portion thereofcomprise a heavy chain sequence of SEQ ID NO: 147. In some embodiments,the anti-Galectin-9 antibodies or antigen-binding portion thereofcomprise a heavy chain sequence of SEQ ID NO: 148. In some embodiments,the anti-Galectin-9 antibodies or antigen-binding portion thereofcomprise a heavy chain sequence of SEQ ID NO: 149. In some embodiments,the anti-Galectin-9 antibodies or antigen-binding portion thereofcomprise a heavy chain sequence of SEQ ID NO: 150. In some embodiments,the anti-Galectin-9 antibodies or antigen-binding portion thereofcomprise a heavy chain sequence of SEQ ID NO: 151. In some embodiments,the anti-Galectin-9 antibodies or antigen-binding portion thereofcomprise a heavy chain sequence of SEQ ID NO: 152. In some embodiments,the anti-Galectin-9 antibodies or antigen-binding portion thereofcomprise a heavy chain sequence of SEQ ID NO: 153. In some embodiments,the anti-Galectin-9 antibodies or antigen-binding portion thereofcomprise a heavy chain sequence of SEQ ID NO: 154. In some embodiments,the anti-Galectin-9 antibodies or antigen-binding portion thereofcomprise a heavy chain sequence of SEQ ID NO: 155. In some embodiments,the anti-Galectin-9 antibodies or antigen-binding portion thereofcomprise a heavy chain sequence of SEQ ID NO: 156. In some embodiments,the anti-Galectin-9 antibodies or antigen-binding portion thereofcomprise a heavy chain sequence of SEQ ID NO: 157. In some embodiments,the anti-Galectin-9 antibodies or antigen-binding portion thereofcomprise a heavy chain sequence of SEQ ID NO: 158. In some embodiments,the anti-Galectin-9 antibodies or antigen-binding portion thereofcomprise a heavy chain sequence of SEQ ID NO: 159. In some embodiments,the anti-Galectin-9 antibodies or antigen-binding portion thereofcomprise a heavy chain sequence of SEQ ID NO: 160. In some embodiments,the anti-Galectin-9 antibodies or antigen-binding portion thereofcomprise a heavy chain sequence of SEQ ID NO: 161. In some embodiments,the anti-Galectin-9 antibodies or antigen-binding portion thereofcomprise a heavy chain sequence of SEQ ID NO: 162. In some embodiments,the anti-Galectin-9 antibodies or antigen-binding portion thereofcomprise a heavy chain sequence of SEQ ID NO: 163. In some embodiments,the anti-Galectin-9 antibodies or antigen-binding portion thereofcomprise a heavy chain sequence of SEQ ID NO: 164. In some embodiments,the anti-Galectin-9 antibodies or antigen-binding portion thereofcomprise a heavy chain sequence of SEQ ID NO: 165. In some embodiments,the anti-Galectin-9 antibodies or antigen-binding portion thereofcomprise a heavy chain sequence of SEQ ID NO: 166. In some embodiments,the anti-Galectin-9 antibodies or antigen-binding portion thereofcomprise a heavy chain sequence of SEQ ID NO: 167. In some embodiments,the anti-Galectin-9 antibodies or antigen-binding portion thereofcomprise a heavy chain sequence of SEQ ID NO: 168.

In some embodiments, the constant region is IgG1 LALA. In someembodiments, the anti-Galectin-9 antibodies or antigen-binding portionthereof comprise a heavy chain sequence of SEQ ID NO: 194. In someembodiments, the anti-Galectin-9 antibodies or antigen-binding portionthereof comprise a heavy chain sequence of SEQ ID NO: 195. In someembodiments, the anti-Galectin-9 antibodies or antigen-binding portionthereof comprise a heavy chain sequence of SEQ ID NO: 196. In someembodiments, the anti-Galectin-9 antibodies or antigen-binding portionthereof comprise a heavy chain sequence of SEQ ID NO: 197. In someembodiments, the anti-Galectin-9 antibodies or antigen-binding portionthereof comprise a heavy chain sequence of SEQ ID NO: 198. In someembodiments, the anti-Galectin-9 antibodies or antigen-binding portionthereof comprise a heavy chain sequence of SEQ ID NO: 199. In someembodiments, the anti-Galectin-9 antibodies or antigen-binding portionthereof comprise a heavy chain sequence of SEQ ID NO: 200. In someembodiments, the anti-Galectin-9 antibodies or antigen-binding portionthereof comprise a heavy chain sequence of SEQ ID NO: 201. In someembodiments, the anti-Galectin-9 antibodies or antigen-binding portionthereof comprise a heavy chain sequence of SEQ ID NO: 202. In someembodiments, the anti-Galectin-9 antibodies or antigen-binding portionthereof comprise a heavy chain sequence of SEQ ID NO: 203. In someembodiments, the anti-Galectin-9 antibodies or antigen-binding portionthereof comprise a heavy chain sequence of SEQ ID NO: 304. In someembodiments, the anti-Galectin-9 antibodies or antigen-binding portionthereof comprise a heavy chain sequence of SEQ ID NO: 205. In someembodiments, the anti-Galectin-9 antibodies or antigen-binding portionthereof comprise a heavy chain sequence of SEQ ID NO: 206. In someembodiments, the anti-Galectin-9 antibodies or antigen-binding portionthereof comprise a heavy chain sequence of SEQ ID NO: 207. In someembodiments, the anti-Galectin-9 antibodies or antigen-binding portionthereof comprise a heavy chain sequence of SEQ ID NO: 208. In someembodiments, the anti-Galectin-9 antibodies or antigen-binding portionthereof comprise a heavy chain sequence of SEQ ID NO: 209. In someembodiments, the anti-Galectin-9 antibodies or antigen-binding portionthereof comprise a heavy chain sequence of SEQ ID NO: 210. In someembodiments, the anti-Galectin-9 antibodies or antigen-binding portionthereof comprise a heavy chain sequence of SEQ ID NO: 211. In someembodiments, the anti-Galectin-9 antibodies or antigen-binding portionthereof comprise a heavy chain sequence of SEQ ID NO: 212. In someembodiments, the anti-Galectin-9 antibodies or antigen-binding portionthereof comprise a heavy chain sequence of SEQ ID NO: 213. In someembodiments, the anti-Galectin-9 antibodies or antigen-binding portionthereof comprise a heavy chain sequence of SEQ ID NO: 214. In someembodiments, the anti-Galectin-9 antibodies or antigen-binding portionthereof comprise a heavy chain sequence of SEQ ID NO: 215. In someembodiments, the anti-Galectin-9 antibodies or antigen-binding portionthereof comprise a heavy chain sequence of SEQ ID NO: 216. In someembodiments, the anti-Galectin-9 antibodies or antigen-binding portionthereof comprise a heavy chain sequence of SEQ ID NO: 217. In someembodiments, the anti-Galectin-9 antibodies or antigen-binding portionthereof comprise a heavy chain sequence of SEQ ID NO: 218. In someembodiments, the anti-Galectin-9 antibodies or antigen-binding portionthereof comprise a heavy chain sequence of SEQ ID NO: 219. In someembodiments, the anti-Galectin-9 antibodies or antigen-binding portionthereof comprise a heavy chain sequence of SEQ ID NO: 220. In someembodiments, the anti-Galectin-9 antibodies or antigen-binding portionthereof comprise a heavy chain sequence of SEQ ID NO: 221.

In some embodiments, the constant region is IgG4. In some embodiments,the anti-Galectin-9 antibodies or antigen-binding portion thereofcomprise a heavy chain sequence of SEQ ID NO: 247. In some embodiments,the anti-Galectin-9 antibodies or antigen-binding portion thereofcomprise a heavy chain sequence of SEQ ID NO: 248. In some embodiments,the anti-Galectin-9 antibodies or antigen-binding portion thereofcomprise a heavy chain sequence of SEQ ID NO: 249. In some embodiments,the anti-Galectin-9 antibodies or antigen-binding portion thereofcomprise a heavy chain sequence of SEQ ID NO: 250. In some embodiments,the anti-Galectin-9 antibodies or antigen-binding portion thereofcomprise a heavy chain sequence of SEQ ID NO: 251. In some embodiments,the anti-Galectin-9 antibodies or antigen-binding portion thereofcomprise a heavy chain sequence of SEQ ID NO: 252. In some embodiments,the anti-Galectin-9 antibodies or antigen-binding portion thereofcomprise a heavy chain sequence of SEQ ID NO: 253. In some embodiments,the anti-Galectin-9 antibodies or antigen-binding portion thereofcomprise a heavy chain sequence of SEQ ID NO: 254. In some embodiments,the anti-Galectin-9 antibodies or antigen-binding portion thereofcomprise a heavy chain sequence of SEQ ID NO: 255. In some embodiments,the anti-Galectin-9 antibodies or antigen-binding portion thereofcomprise a heavy chain sequence of SEQ ID NO: 256. In some embodiments,the anti-Galectin-9 antibodies or antigen-binding portion thereofcomprise a heavy chain sequence of SEQ ID NO: 257. In some embodiments,the anti-Galectin-9 antibodies or antigen-binding portion thereofcomprise a heavy chain sequence of SEQ ID NO: 258. In some embodiments,the anti-Galectin-9 antibodies or antigen-binding portion thereofcomprise a heavy chain sequence of SEQ ID NO: 259. In some embodiments,the anti-Galectin-9 antibodies or antigen-binding portion thereofcomprise a heavy chain sequence of SEQ ID NO: 260. In some embodiments,the anti-Galectin-9 antibodies or antigen-binding portion thereofcomprise a heavy chain sequence of SEQ ID NO: 261. In some embodiments,the anti-Galectin-9 antibodies or antigen-binding portion thereofcomprise a heavy chain sequence of SEQ ID NO: 262. In some embodiments,the anti-Galectin-9 antibodies or antigen-binding portion thereofcomprise a heavy chain sequence of SEQ ID NO: 263. In some embodiments,the anti-Galectin-9 antibodies or antigen-binding portion thereofcomprise a heavy chain sequence of SEQ ID NO: 264. In some embodiments,the anti-Galectin-9 antibodies or antigen-binding portion thereofcomprise a heavy chain sequence of SEQ ID NO: 265. In some embodiments,the anti-Galectin-9 antibodies or antigen-binding portion thereofcomprise a heavy chain sequence of SEQ ID NO: 266. In some embodiments,the anti-Galectin-9 antibodies or antigen-binding portion thereofcomprise a heavy chain sequence of SEQ ID NO: 267. In some embodiments,the anti-Galectin-9 antibodies or antigen-binding portion thereofcomprise a heavy chain sequence of SEQ ID NO: 268. In some embodiments,the anti-Galectin-9 antibodies or antigen-binding portion thereofcomprise a heavy chain sequence of SEQ ID NO: 269. In some embodiments,the anti-Galectin-9 antibodies or antigen-binding portion thereofcomprise a heavy chain sequence of SEQ ID NO: 270. In some embodiments,the anti-Galectin-9 antibodies or antigen-binding portion thereofcomprise a heavy chain sequence of SEQ ID NO: 271. In some embodiments,the anti-Galectin-9 antibodies or antigen-binding portion thereofcomprise a heavy chain sequence of SEQ ID NO: 272. In some embodiments,the anti-Galectin-9 antibodies or antigen-binding portion thereofcomprise a heavy chain sequence of SEQ ID NO: 273. In some embodiments,the anti-Galectin-9 antibodies or antigen-binding portion thereofcomprise a heavy chain sequence of SEQ ID NO: 274.

In some embodiments, the constant region is IgG4 mut. In someembodiments, the anti-Galectin-9 antibodies or antigen-binding portionthereof comprise a heavy chain sequence of SEQ ID NO: 300. In someembodiments, the anti-Galectin-9 antibodies or antigen-binding portionthereof comprise a heavy chain sequence of SEQ ID NO: 301. In someembodiments, the anti-Galectin-9 antibodies or antigen-binding portionthereof comprise a heavy chain sequence of SEQ ID NO: 302. In someembodiments, the anti-Galectin-9 antibodies or antigen-binding portionthereof comprise a heavy chain sequence of SEQ ID NO: 303. In someembodiments, the anti-Galectin-9 antibodies or antigen-binding portionthereof comprise a heavy chain sequence of SEQ ID NO: 304. In someembodiments, the anti-Galectin-9 antibodies or antigen-binding portionthereof comprise a heavy chain sequence of SEQ ID NO: 305. In someembodiments, the anti-Galectin-9 antibodies or antigen-binding portionthereof comprise a heavy chain sequence of SEQ ID NO: 306. In someembodiments, the anti-Galectin-9 antibodies or antigen-binding portionthereof comprise a heavy chain sequence of SEQ ID NO: 307. In someembodiments, the anti-Galectin-9 antibodies or antigen-binding portionthereof comprise a heavy chain sequence of SEQ ID NO: 308. In someembodiments, the anti-Galectin-9 antibodies or antigen-binding portionthereof comprise a heavy chain sequence of SEQ ID NO: 309. In someembodiments, the anti-Galectin-9 antibodies or antigen-binding portionthereof comprise a heavy chain sequence of SEQ ID NO: 310. In someembodiments, the anti-Galectin-9 antibodies or antigen-binding portionthereof comprise a heavy chain sequence of SEQ ID NO: 311. In someembodiments, the anti-Galectin-9 antibodies or antigen-binding portionthereof comprise a heavy chain sequence of SEQ ID NO: 312. In someembodiments, the anti-Galectin-9 antibodies or antigen-binding portionthereof comprise a heavy chain sequence of SEQ ID NO: 313. In someembodiments, the anti-Galectin-9 antibodies or antigen-binding portionthereof comprise a heavy chain sequence of SEQ ID NO: 314. In someembodiments, the anti-Galectin-9 antibodies or antigen-binding portionthereof comprise a heavy chain sequence of SEQ ID NO: 315. In someembodiments, the anti-Galectin-9 antibodies or antigen-binding portionthereof comprise a heavy chain sequence of SEQ ID NO: 316. In someembodiments, the anti-Galectin-9 antibodies or antigen-binding portionthereof comprise a heavy chain sequence of SEQ ID NO: 317. In someembodiments, the anti-Galectin-9 antibodies or antigen-binding portionthereof comprise a heavy chain sequence of SEQ ID NO: 318. In someembodiments, the anti-Galectin-9 antibodies or antigen-binding portionthereof comprise a heavy chain sequence of SEQ ID NO: 319. In someembodiments, the anti-Galectin-9 antibodies or antigen-binding portionthereof comprise a heavy chain sequence of SEQ ID NO: 320. In someembodiments, the anti-Galectin-9 antibodies or antigen-binding portionthereof comprise a heavy chain sequence of SEQ ID NO: 321. In someembodiments, the anti-Galectin-9 antibodies or antigen-binding portionthereof comprise a heavy chain sequence of SEQ ID NO: 322. In someembodiments, the anti-Galectin-9 antibodies or antigen-binding portionthereof comprise a heavy chain sequence of SEQ ID NO: 323. In someembodiments, the anti-Galectin-9 antibodies or antigen-binding portionthereof comprise a heavy chain sequence of SEQ ID NO: 324. In someembodiments, the anti-Galectin-9 antibodies or antigen-binding portionthereof comprise a heavy chain sequence of SEQ ID NO: 325. In someembodiments, the anti-Galectin-9 antibodies or antigen-binding portionthereof comprise a heavy chain sequence of SEQ ID NO: 326.

In some embodiments, heavy chains of anti-Galectin-9 antibodies comprisean amino acid sequence that has at least 80% (e.g., 85%, 90%, 95%, 96%,97%, 98%, 99% and any increment therein) sequence identity to the heavychain sequence set forth in SEQ ID NO: 157 (or its variable region). Insome embodiments, heavy chains of anti-Galectin-9 antibodies consist ofan amino acid sequence that has at least 80% (e.g., 85%, 90%, 95%, 96%,97%, 98%, 99% and any increment therein) sequence identity to the heavychain sequence set forth in SEQ ID NO: 157. In some embodiments, heavychains of anti-Galectin-9 antibodies comprise an amino acid sequence setforth in SEQ ID NO: 157 (or its variable region). In some embodiments,heavy chains of anti-Galectin-9 antibodies consist of an amino acidsequence set forth in SEQ ID NO: 157.

In some embodiments, heavy chains of anti-Galectin-9 antibodies comprisean amino acid sequence that has at least 80% (e.g., 85%, 90%, 95%, 96%,97%, 98%, 99% and any increment therein) sequence identity to the heavychain sequence set forth in SEQ ID NO: 210 (or its variable region). Insome embodiments, heavy chains of anti-Galectin-9 antibodies consist ofan amino acid sequence that has at least 80% (e.g., 85%, 90%, 95%, 96%,97%, 98%, 99% and any increment therein) sequence identity to the heavychain sequence set forth in SEQ ID NO: 210. In some embodiments, heavychains of anti-Galectin-9 antibodies comprise an amino acid sequence setforth in SEQ ID NO: 210 (or its variable region). In some embodiments,heavy chains of anti-Galectin-9 antibodies consist of an amino acidsequence set forth in SEQ ID NO: 210. In some embodiments, heavy chainsof anti-Galectin-9 antibodies comprise an amino acid sequence that hasat least 80% (e.g., 85%, 90%, 95%, 96%, 97%, 98%, 99% and any incrementtherein) sequence identity to the heavy chain sequence set forth in SEQID NO: 263 (or its variable region). In some embodiments, heavy chainsof anti-Galectin-9 antibodies consist of an amino acid sequence that hasat least 80% (e.g., 85%, 90%, 95%, 96%, 97%, 98%, 99% and any incrementtherein) sequence identity to the heavy chain sequence set forth in SEQID NO: 263.

In some embodiments, heavy chains of anti-Galectin-9 antibodies comprisean amino acid sequence set forth in SEQ ID NO: 263 (or its variableregion). In some embodiments, heavy chains of anti-Galectin-9 antibodiesconsist of an amino acid sequence set forth in SEQ ID NO: 263.

In some embodiments, heavy chains of anti-Galectin-9 antibodies comprisean amino acid sequence that has at least 80% (e.g., 85%, 90%, 95%, 96%,97%, 98%, 99% and any increment therein) sequence identity to the heavychain sequence set forth in SEQ ID NO: 316 (or its variable region). Insome embodiments, heavy chains of anti-Galectin-9 antibodies consist ofan amino acid sequence that has at least 80% (e.g., 85%, 90%, 95%, 96%,97%, 98%, 99% and any increment therein) sequence identity to the heavychain sequence set forth in SEQ ID NO: 316. In some embodiments, heavychains of anti-Galectin-9 antibodies comprise an amino acid sequence setforth in SEQ ID NO: 316 (or its variable region). In some embodiments,heavy chains of anti-Galectin-9 antibodies consist of an amino acidsequence set forth in SEQ ID NO: 316.

Clone 9.2 Derived Heavy and Light Chains

In some embodiments, the amino acid sequences of exemplary anti-Galectinantibody light chains correspond to SEQ ID NO: 99-108, and the aminoacid sequences of the exemplary anti-Galectin antibody heavy chainscorrespond to SEQ ID NO: 141-168; 194-221; 249-274; 300-327.

In some embodiments, the anti-Galectin-9 antibodies or antigen-bindingportion thereof comprise a light chain having the sequence of SEQ ID NO:99 and a heavy chain having a sequence selected from SEQ ID NO: 141,194, 247, or 300.

In some embodiments, the anti-Galectin-9 antibodies or antigen-bindingportion thereof comprise a light chain having the sequence of SEQ ID NO:91 and a heavy chain having a sequence selected from SEQ ID NO: 142,195, 248, or 301.

In some embodiments, the anti-Galectin-9 antibodies or antigen-bindingportion thereof comprise a light chain having the sequence of SEQ ID NO:91 and a heavy chain having a sequence selected from SEQ ID NO: 143,196, 249, or 302.

In some embodiments, the anti-Galectin-9 antibodies or antigen-bindingportion thereof comprise a light chain having the sequence of SEQ ID NO:91 and a heavy chain having a sequence selected from SEQ ID NO: 144,197, 250, or 303.

In some embodiments, the anti-Galectin-9 antibodies or antigen-bindingportion thereof comprise a light chain having the sequence of SEQ ID NO:100 and a heavy chain having a sequence selected from SEQ ID NO: 145,198, 251, or 304.

In some embodiments, the anti-Galectin-9 antibodies or antigen-bindingportion thereof comprise a light chain having the sequence of SEQ ID NO:101 and a heavy chain having a sequence selected from SEQ ID NO: 146,199, 252, or 305.

In some embodiments, the anti-Galectin-9 antibodies or antigen-bindingportion thereof comprise a light chain having the sequence of SEQ ID NO:102 and a heavy chain having a sequence selected from SEQ ID NO: 147,200, 253, or 306.

In some embodiments, the anti-Galectin-9 antibodies or antigen-bindingportion thereof comprise a light chain having the sequence of SEQ ID NO:103 and a heavy chain having a sequence selected from SEQ ID NO: 148,201, 254, or 307.

In some embodiments, the anti-Galectin-9 antibodies or antigen-bindingportion thereof comprise a light chain having the sequence of SEQ ID NO:104 and a heavy chain having a sequence selected from SEQ ID NO: 149,202, 255, or 308.

In some embodiments, the anti-Galectin-9 antibodies or antigen-bindingportion thereof comprise a light chain having the sequence of SEQ ID NO:105 and a heavy chain having a sequence selected from SEQ ID NO: 150,203, 256, or 309.

In some embodiments, the anti-Galectin-9 antibodies or antigen-bindingportion thereof comprise a light chain having the sequence of SEQ ID NO:106 and a heavy chain having a sequence selected from SEQ ID NO: 151,204, 257, or 310.

In some embodiments, the anti-Galectin-9 antibodies or antigen-bindingportion thereof comprise a light chain having the sequence of SEQ ID NO:107 and a heavy chain having a sequence selected from SEQ ID NO: 152,205, 258, or 311.

In some embodiments, the anti-Galectin-9 antibodies or antigen-bindingportion thereof comprise a light chain having the sequence of SEQ ID NO:99 and a heavy chain having a sequence selected from SEQ ID NO: 153,206, 259, or 312.

In some embodiments, the anti-Galectin-9 antibodies or antigen-bindingportion thereof comprise a light chain having the sequence of SEQ ID NO:91 and a heavy chain having a sequence selected from SEQ ID NO: 154,207, 260, or 313.

In some embodiments, the anti-Galectin-9 antibodies or antigen-bindingportion thereof comprise a light chain having the sequence of SEQ ID NO:100 and a heavy chain having a sequence selected from SEQ ID NO: 155,208, 261, or 314.

In some embodiments, the anti-Galectin-9 antibodies or antigen-bindingportion thereof comprise a light chain having the sequence of SEQ ID NO:91 and a heavy chain having a sequence selected from SEQ ID NO: 156,209, 262, or 315.

In some embodiments, the anti-Galectin-9 antibodies or antigen-bindingportion thereof comprise a light chain having the sequence of SEQ ID NO:108 and a heavy chain having a sequence selected from SEQ ID NO: 157,210, 263, or 316.

In some embodiments, the anti-Galectin-9 antibodies or antigen-bindingportion thereof comprise a light chain having the sequence of SEQ ID NO:108 and a heavy chain having a sequence selected from SEQ ID NO: 158,211, 264, or 317.

In some embodiments, the anti-Galectin-9 antibodies or antigen-bindingportion thereof comprise a light chain having the sequence of SEQ ID NO:91 and a heavy chain having a sequence selected from SEQ ID NO: 159,212, 265, or 318.

In some embodiments, the anti-Galectin-9 antibodies or antigen-bindingportion thereof comprise a light chain having the sequence of SEQ ID NO:109 and a heavy chain having a sequence selected from SEQ ID NO: 160,213, 266, or 319.

In some embodiments, the anti-Galectin-9 antibodies or antigen-bindingportion thereof comprise a light chain having the sequence of SEQ ID NO:108 and a heavy chain having a sequence selected from SEQ ID NO: 161,214, 267, or 320.

In some embodiments, the anti-Galectin-9 antibodies or antigen-bindingportion thereof comprise a light chain having the sequence of SEQ ID NO:110 and a heavy chain having a sequence selected from SEQ ID NO: 162,215, 268, or 321.

In some embodiments, the anti-Galectin-9 antibodies or antigen-bindingportion thereof comprise a light chain having the sequence of SEQ ID NO:111 and a heavy chain having a sequence selected from SEQ ID NO: 163,216, 269, or 322.

In some embodiments, the anti-Galectin-9 antibodies or antigen-bindingportion thereof comprise a light chain having the sequence of SEQ ID NO:112 and a heavy chain having a sequence selected from SEQ ID NO: 164,217, 270, or 323.

In some embodiments, the anti-Galectin-9 antibodies or antigen-bindingportion thereof comprise a light chain having the sequence of SEQ ID NO:113 and a heavy chain having a sequence selected from SEQ ID NO: 165,218, 271, or 324.

In some embodiments, the anti-Galectin-9 antibodies or antigen-bindingportion thereof comprise a light chain having the sequence of SEQ ID NO:114 and a heavy chain having a sequence selected from SEQ ID NO: 166,219, 272, or 325.

In some embodiments, the anti-Galectin-9 antibodies or antigen-bindingportion thereof comprise a light chain having the sequence of SEQ ID NO:115 and a heavy chain having a sequence selected from SEQ ID NO: 167,220, 273, or 326.

In some embodiments, the anti-Galectin-9 antibodies or antigen-bindingportion thereof comprise a light chain having the sequence of SEQ ID NO:108 and a heavy chain having a sequence selected from SEQ ID NO: 168,221, 274, or 327.

In some embodiments, the anti-Galectin-9 antibodies or antigen-bindingportion thereof comprise a light chain sequence of SEQ ID NO: 108 andcomprise a heavy chain sequence of SEQ ID NO: 157. In some embodiments,light chains of anti-Galectin-9 antibodies comprise an amino acidsequence that has at least 80% (e.g., 85%, 90%, 95%, 96%, 97%, 98%, 99%and any increment therein) sequence identity to the light chain sequenceset forth in SEQ ID NO: 108 (or their variable regions), and heavychains of anti-Galectin-9 antibodies comprise an amino acid sequencethat has at least 80% (e.g., 85%, 90%, 95%, 96%, 97%, 98%, 99% and anyincrement therein) sequence identity to the heavy chain sequence setforth in SEQ ID NO: 157 (or its variable region). In some embodiments,light chains of anti-Galectin-9 antibodies consist of an amino acidsequence that has at least 80% (e.g., 85%, 90%, 95%, 96%, 97%, 98%, 99%and any increment therein) sequence identity to the light chain sequenceset forth in SEQ ID NO: 108 and heavy chains of anti-Galectin-9antibodies consist of an amino acid sequence that has at least 80%(e.g., 85%, 90%, 95%, 96%, 97%, 98%, 99% and any increment therein)sequence identity to the heavy chain sequence set forth in SEQ ID NO:157. In some embodiments, light chains of anti-Galectin-9 antibodiescomprise an amino acid sequence set forth in SEQ ID NO: 108 (or theirvariable regions) heavy chains of anti-Galectin-9 antibodies comprise anamino acid sequence set forth in SEQ ID NO: 157 (or its variableregion). In some embodiments, light chains of anti-Galectin-9 antibodiesconsist of the amino acid sequence set forth in SEQ ID NO: 108 and heavychains of anti-Galectin-9 antibodies consist of an amino acid sequenceset forth in SEQ ID NO: 157.

In some embodiments, the anti-Galectin-9 antibodies or antigen-bindingportion thereof comprise a light chain sequence of SEQ ID NO: 108 andcomprise a heavy chain sequence of SEQ ID NO: 210. In some embodiments,light chains of anti-Galectin-9 antibodies comprise an amino acidsequence that has at least 80% (e.g., 85%, 90%, 95%, 96%, 97%, 98%, 99%and any increment therein) sequence identity to the light chain sequenceset forth in SEQ ID NO: 108 (or their variable regions) and heavy chainsof anti-Galectin-9 antibodies comprise an amino acid sequence that hasat least 80% (e.g., 85%, 90%, 95%, 96%, 97%, 98%, 99% and any incrementtherein) sequence identity to the heavy chain sequence set forth in SEQID NO: 210 (or its variable region) and heavy chains of anti-Galectin-9antibodies consist of an amino acid sequence that has at least 80%(e.g., 85%, 90%, 95%, 96%, 97%, 98%, 99% and any increment therein)sequence identity to the heavy chain sequence set forth in SEQ ID NO:210. In some embodiments, light chains of anti-Galectin-9 antibodiesconsist of an amino acid sequence that has at least 80% (e.g., 85%, 90%,95%, 96%, 97%, 98%, 99% and any increment therein) sequence identity tothe light chain sequence set forth in SEQ ID NO: 108. In someembodiments, light chains of anti-Galectin-9 antibodies comprise anamino acid sequence set forth in SEQ ID NO: 108 (or their variableregions) and heavy chains of anti-Galectin-9 antibodies comprise anamino acid sequence set forth in SEQ ID NO: 210 (or its variableregion). In some embodiments, light chains of anti-Galectin-9 antibodiesconsist of the amino acid sequence set forth in SEQ ID NO: 108 and heavychains of anti-Galectin-9 antibodies comprise an amino acid sequence setforth in SEQ ID NO: 210.

In some embodiments, the anti-Galectin-9 antibodies or antigen-bindingportion thereof comprise a light chain sequence of SEQ ID NO: 108 andcomprise a heavy chain sequence of SEQ ID NO: 263. In some embodiments,light chains of anti-Galectin-9 antibodies comprise an amino acidsequence that has at least 80% (e.g., 85%, 90%, 95%, 96%, 97%, 98%, 99%and any increment therein) sequence identity to the light chain sequenceset forth in SEQ ID NO: 108 (or their variable regions) and heavy chainsof anti-Galectin-9 antibodies comprise an amino acid sequence that hasat least 80% (e.g., 85%, 90%, 95%, 96%, 97%, 98%, 99% and any incrementtherein) sequence identity to the heavy chain sequence set forth in SEQID NO: 263 (or its variable region). In some embodiments, light chainsof anti-Galectin-9 antibodies consist of an amino acid sequence that hasat least 80% (e.g., 85%, 90%, 95%, 96%, 97%, 98%, 99% and any incrementtherein) sequence identity to the light chain sequence set forth in SEQID NO: 108 and heavy chains of anti-Galectin-9 antibodies consist of anamino acid sequence that has at least 80% (e.g., 85%, 90%, 95%, 96%,97%, 98%, 99% and any increment therein) sequence identity to the heavychain sequence set forth in SEQ ID NO: 263. In some embodiments, lightchains of anti-Galectin-9 antibodies comprise an amino acid sequence setforth in SEQ ID NO: 108 (or their variable regions) and heavy chains ofanti-Galectin-9 antibodies comprise an amino acid sequence set forth inSEQ ID NO: 263 (or its variable region). In some embodiments, lightchains of anti-Galectin-9 antibodies consist of the amino acid sequenceset forth in SEQ ID NO: 108 and heavy chains of anti-Galectin-9antibodies consist of an amino acid sequence set forth in SEQ ID NO:263.

In some embodiments, the anti-Galectin-9 antibodies or antigen-bindingportion thereof comprise a light chain sequence of SEQ ID NO: 108 andcomprise a heavy chain sequence of SEQ ID NO: 316. In some embodiments,light chains of anti-Galectin-9 antibodies comprise an amino acidsequence that has at least 80% (e.g., 85%, 90%, 95%, 96%, 97%, 98%, 99%and any increment therein) sequence identity to the light chain sequenceset forth in SEQ ID NO: 108 (or their variable regions) and heavy chainsof anti-Galectin-9 antibodies comprise an amino acid sequence that hasat least 80% (e.g., 85%, 90%, 95%, 96%, 97%, 98%, 99% and any incrementtherein) sequence identity to the heavy chain sequence set forth in SEQID NO: 316 (or its variable region). In some embodiments, light chainsof anti-Galectin-9 antibodies consist of an amino acid sequence that hasat least 80% (e.g., 85%, 90%, 95%, 96%, 97%, 98%, 99% and any incrementtherein) sequence identity to the light chain sequence set forth in SEQID NO: 108 and heavy chains of anti-Galectin-9 antibodies consist of anamino acid sequence that has at least 80% (e.g., 85%, 90%, 95%, 96%,97%, 98%, 99% and any increment therein) sequence identity to the heavychain sequence set forth in SEQ ID NO: 316. In some embodiments, lightchains of anti-Galectin-9 antibodies comprise an amino acid sequence setforth in SEQ ID NO: 108 (or their variable regions) and heavy chains ofanti-Galectin-9 antibodies comprise an amino acid sequence set forth inSEQ ID NO: 316. In some embodiments, light chains of anti-Galectin-9antibodies consist of the amino acid sequence set forth in SEQ ID NO:108 and heavy chains of anti-Galectin-9 antibodies consist of an aminoacid sequence set forth in SEQ ID NO: 316.

In some embodiments, the anti-Galectin-9 antibody comprises an IgG1heavy chain having the sequence of SEQ ID NO: 157 and a light chainhaving the sequence of SEQ ID NO: 108. In some embodiments, theanti-Galectin-9 antibody comprises an IgG1 heavy chain having thesequence of SEQ ID NO: 210 and a light chain having the sequence of SEQID NO: 108. In some embodiments, the anti-Galectin-9 antibody comprisesan IgG4 heavy chain having the sequence of SEQ ID NO: 263 and a lightchain having the sequence of SEQ ID NO: 108. In some embodiments, theanti-Galectin-9 antibody comprises an IgG4 heavy chain having thesequence of SEQ ID NO: 316 and a light chain having the sequence of SEQID NO: 108. In some embodiments, the anti-Galectin-9 antibody comprisesan IgG1 heavy chain having the sequence of SEQ ID NO: 136 and a lightchain having the sequence of SEQ ID NO: 108. In some embodiments, theanti-Galectin-9 antibody comprises an IgG1 heavy chain having thesequence of SEQ ID NO: 189 and a light chain having the sequence of SEQID NO: 108. In some embodiments, the anti-Galectin-9 antibody comprisesan IgG4 heavy chain having the sequence of SEQ ID NO: 242 and a lightchain having the sequence of SEQ ID NO: 108. In some embodiments, theanti-Galectin-9 antibody comprises an IgG4 heavy chain having thesequence of SEQ ID NO: 295 and a light chain having the sequence of SEQID NO: 108.

Antibody heavy and light chain constant regions are well known in theart, e.g., those provided in the IMGT database (www.imgt.org) or atwww.vbase2.org/vbstat.php., both of which are incorporated by referenceherein.

Preparation of Anti-Galectin-9 Antibodies

Antibodies capable of binding Galectin-9 as described herein can be madeby any method known in the art. See, for example, Harlow and Lane,(1998) Antibodies: A Laboratory Manual, Cold Spring Harbor Laboratory,New York.

In some embodiments, antibodies specific to a target antigen (e.g.,Galectin-9 or a CRD thereof) are made by conventional hybridomatechnology. The full-length target antigen or a fragment thereof,optionally coupled to a carrier protein such as KLH, can be used toimmunize a host animal for generating antibodies binding to thatantigen. The route and schedule of immunization of the host animal aregenerally in keeping with established and conventional techniques forantibody stimulation and production, as further described herein.General techniques for production of mouse, humanized, and humanantibodies are known in the art and are described herein. It iscontemplated that any mammalian subject including humans or antibodyproducing cells therefrom can be manipulated to serve as the basis forproduction of mammalian, including human hybridoma cell lines.Typically, the host animal is inoculated intraperitoneally,intramuscularly, orally, subcutaneously, intraplantar, and/orintradermally with an amount of immunogen, including as describedherein.

Hybridomas can be prepared from the lymphocytes and immortalized myelomacells using the general somatic cell hybridization technique of Kohler,B. and Milstein, C. (1975) Nature 256:495-497 or as modified by Buck, D.W., et al., In Vitro, 18:377-381 (1982). Available myeloma lines,including, but not limited to, X63-Ag8.653 and those from the SalkInstitute, Cell Distribution Center, San Diego, Calif., USA, may be usedin the hybridization. Generally, the technique involves fusing myelomacells and lymphoid cells using a fusogen such as polyethylene glycol, orby electrical means well known to those skilled in the art. After thefusion, the cells are separated from the fusion medium and grown in aselective growth medium, such as hypoxanthine-aminopterin-thymidine(HAT) medium, to eliminate unhybridized parent cells. Any of the mediadescribed herein, supplemented with or without serum, can be used forculturing hybridomas that secrete monoclonal antibodies. As anotheralternative to the cell fusion technique, EBV immortalized B cells maybe used to produce the anti-Galectin-9 monoclonal antibodies describedherein. The hybridomas are expanded and subcloned, if desired, andsupernatants are assayed for anti-immunogen activity by conventionalimmunoassay procedures (e.g., radioimmunoassay, enzyme immunoassay, orfluorescence immunoassay).

Hybridomas that may be used as source of antibodies encompass allderivatives, progeny cells of the parent hybridomas that producemonoclonal antibodies capable of interfering with the Galectin-9activity. Hybridomas that produce such antibodies may be grown in vitroor in vivo using known procedures. The monoclonal antibodies may beisolated from the culture media or body fluids, by conventionalimmunoglobulin purification procedures such as ammonium sulfateprecipitation, gel electrophoresis, dialysis, chromatography, andultrafiltration, if desired. Undesired activity if present, can beremoved, for example, by running the preparation over adsorbents made ofthe immunogen attached to a solid phase and eluting or releasing thedesired antibodies off the immunogen. Immunization of a host animal witha target antigen or a fragment containing the target amino acid sequenceconjugated to a protein that is immunogenic in the species to beimmunized, e.g., keyhole limpet hemocyanin, serum albumin, bovinethyroglobulin, or soybean trypsin inhibitor using a bifunctional orderivatizing agent, for example maleimidobenzoyl sulfosuccinimide ester(conjugation through cysteine residues), N-hydroxysuccinimide (throughlysine residues), glutaraldehyde, succinic anhydride, SOCl, or R1N═C═NR,where R and R1 are different alkyl groups, can yield a population ofantibodies (e.g., monoclonal antibodies).

If desired, an antibody (monoclonal or polyclonal) of interest (e.g.,produced by a hybridoma) may be sequenced and the polynucleotidesequence may then be cloned into a vector for expression or propagation.The sequence encoding the antibody of interest may be maintained invector in a host cell and the host cell can then be expanded and frozenfor future use. In an alternative, the polynucleotide sequence may beused for genetic manipulation to “humanize” the antibody or to improvethe affinity (affinity maturation), or other characteristics of theantibody. For example, the constant region may be engineered to moreresemble human constant regions to avoid immune response if the antibodyis used in clinical trials and treatments in humans. It may be desirableto genetically manipulate the antibody sequence to obtain greateraffinity to the target antigen and greater efficacy in inhibiting theactivity of Galectin-9. It will be apparent to one of skill in the artthat one or more polynucleotide changes can be made to the antibody andstill maintain its binding specificity to the target antigen.

In other embodiments, fully human antibodies are obtained usingcommercially available mice that have been engineered to expressspecific human immunoglobulin proteins. Transgenic animals that aredesigned to produce a more desirable (e.g., fully human antibodies) ormore robust immune response may also be used for generation of humanizedor human antibodies. Examples of such technology are Xenomouse® fromAmgen, Inc. (Fremont, Calif.) and HuMAb-Mouse® and TC Mouse™ fromMedarex, Inc. (Princeton, N.J.). In other embodiments, antibodies aremade recombinantly by phage display or yeast technology. See, forexample, U.S. Pat. Nos. 5,565,332; 5,580,717; 5,733,743; and 6,265,150;and Winter et al., (1994) Annu. Rev. Immunol. 12:433-455. In alternateembodiments, phage display technology (McCafferty et al., (1990) Nature348:552-553) is used to produce human antibodies and antibody fragmentsin vitro, from immunoglobulin variable (V) domain gene repertoires fromunimmunized donors.

In alternate embodiments, antibodies capable of binding to the targetantigens as described herein are isolated from a suitable antibodylibrary. Antibody libraries, which contain a plurality of antibodycomponents, can be used to identify antibodies that bind to a specifictarget antigen (e.g., the CRD1 or CRD2 of Galectin-9 in this case)following routine selection processes as known in the art. In theselection process, an antibody library can be probed with the targetantigen or a fragment thereof and members of the library that arecapable of binding to the target antigen can be isolated, typically byretention on a support. Such screening process may be performed bymultiple rounds (e.g., including both positive and negative selections)to enrich the pool of antibodies capable of binding to the targetantigen. Individual clones of the enriched pool can then be isolated andfurther characterized to identify those having desired binding activityand biological activity. Sequences of the heavy chain and light chainvariable domains can also be determined via conventional methodology.There are a number of routine methods known in the art to identify andisolate antibodies capable of binding to the target antigens describedherein, including phage display, yeast display, ribosomal display, ormammalian display technology.

As an example, phage displays typically use a covalent linkage to bindthe protein (e.g., antibody) component to a bacteriophage coat protein.The linkage results from translation of a nucleic acid encoding theantibody component fused to the coat protein. The linkage can include aflexible peptide linker, a protease site, or an amino acid incorporatedas a result of suppression of a stop codon. Phage display is described,for example, in U.S. Pat. No. 5,223,409; Smith (1985) Science228:1315-1317; WO 92/18619; WO 91/17271; WO 92/20791; WO 92/15679; WO93/01288; WO 92/01047; WO 92/09690; WO 90/02809; de Haard et al. (1999)J. Biol. Chem 274:18218-30; Hoogenboom et al. (1998) Immunotechnology4:1-20; Hoogenboom et al. (2000) Immunol Today 2:371-8 and Hoet et al.(2005) Nat Biotechnol. 23(3)344-8. Additional suitable methods aredescribed in Miller et al., PloS One, 2012, 7, e43746; Fellouse et al.,J Mol Biol, 2007, 373, 924-940. Bacteriophage displaying the proteincomponent can be grown and harvested using standard phage preparatorymethods, e.g. PEG precipitation from growth media. After selection ofindividual display phages, the nucleic acid encoding the selectedprotein components can be isolated from cells infected with the selectedphages or from the phage themselves, after amplification. Individualcolonies or plaques can be selected, and then the nucleic acid may beisolated and sequenced.

Other display formats include cell-based display (see, e.g., WO03/029456), protein-nucleic acid fusions (see, e.g., U.S. Pat. No.6,207,446), ribosome display (See, e.g., Mattheakis et al. (1994) Proc.Natl. Acad. Sci. USA 91:9022 and Hanes et al. (2000) Nat Biotechnol.18:1287-92; Hanes et al. (2000) Methods Enzymol. 328:404-30; andSchaffitzel et al. (1999) J Immunol Methods. 231(1-2):119-35), and E.coli periplasmic display (J Immunol Methods. 2005 Nov. 22; PMID:16337958).

After display library members are isolated for binding to the targetantigen, each isolated library member can be also tested for its abilityto bind to a non-target molecule to evaluate its binding specificity.Examples of non-target molecules include streptavidin on magnetic beads,blocking agents such as bovine serum albumin, non-fat bovine milk, soyprotein, any capturing or target immobilizing monoclonal antibody, ornon-transfected cells which do not express the target. A high-throughputELISA screen can be used to obtain the data, for example. The ELISAscreen can also be used to obtain quantitative data for binding of eachlibrary member to the target as well as for cross species reactivity torelated targets or subunits of the target antigen and also underdifferent condition such as pH 6 or pH 7.5. The non-target and targetbinding data are compared (e.g., using a computer and software) toidentify library members that specifically bind to the target.

After selecting candidate library members that bind to a target, eachcandidate library member can be further analyzed, e.g., to furthercharacterize its binding properties for the target, e.g., Galectin-9.Each candidate library member can be subjected to one or more secondaryscreening assays. The assay can be for a binding property, a catalyticproperty, an inhibitory property, a physiological property (e.g.,cytotoxicity, renal clearance, immunogenicity), a structural property(e.g., stability, conformation, oligomerization state) or anotherfunctional property. The same assay can be used repeatedly, but withvarying conditions, e.g., to determine pH, ionic, or thermalsensitivities.

As appropriate, the assays can use a display library member directly, arecombinant polypeptide produced from the nucleic acid encoding theselected polypeptide, or a synthetic peptide synthesized based on thesequence of the selected polypeptide. In the case of selected Fabs, theFabs can be evaluated or can be modified and produced as intact IgGproteins. Exemplary assays for binding properties are described below.

Binding proteins can also be evaluated using an ELISA assay. Forexample, each protein is contacted to a microtitre plate whose bottomsurface has been coated with the target, e.g., a limiting amount of thetarget. The plate is washed with buffer to remove non-specifically boundpolypeptides. Then the amount of the binding protein bound to the targeton the plate is determined by probing the plate with an antibody thatcan recognize the binding protein, e.g., a tag or constant portion ofthe binding protein. The antibody is linked to a detection system (e.g.,an enzyme such as alkaline phosphatase or horse radish peroxidase (HRP)which produces a colorimetric product when appropriate substrates areprovided).

Alternatively, the ability of a binding protein described herein to binda target antigen can be analyzed using a homogenous assay, i.e., afterall components of the assay are added, additional fluid manipulationsare not required. For example, fluorescence resonance energy transfer(FRET) can be used as a homogenous assay (see, for example, Lakowicz etal., U.S. Pat. No. 5,631,169; Stavrianopoulos, et al., U.S. Pat. No.4,868,103). A fluorophore label on the first molecule (e.g., themolecule identified in the fraction) is selected such that its emittedfluorescent energy can be absorbed by a fluorescent label on a secondmolecule (e.g., the target) if the second molecule is in proximity tothe first molecule. The fluorescent label on the second moleculefluoresces when it absorbs to the transferred energy. Since theefficiency of energy transfer between the labels is related to thedistance separating the molecules, the spatial relationship between themolecules can be assessed. In a situation in which binding occursbetween the molecules, the fluorescent emission of the ‘acceptor’molecule label in the assay should be maximal. A binding event that isconfigured for monitoring by FRET can be conveniently measured throughstandard fluorometric detection means, e.g., using a fluorimeter. Bytitrating the amount of the first or second binding molecule, a bindingcurve can be generated to estimate the equilibrium binding constant.

Surface plasmon resonance (SPR) can be used to analyze the interactionof a binding protein and a target antigen. SPR or BiomolecularInteraction Analysis (BIA) detects biospecific interactions in realtime, without labeling any of the interactants. Changes in the mass atthe binding surface (indicative of a binding event) of the BIA chipresult in alterations of the refractive index of light near the surface(the optical phenomenon of SPR). The changes in the refractivitygenerate a detectable signal, which are measured as an indication ofreal-time reactions between biological molecules. Methods for using SPRare described, for example, in U.S. Pat. No. 5,641,640; Raether, 1988,Surface Plasmons Springer Verlag; Sjolander and Urbaniczky, 1991, Anal.Chem. 63:2338-2345; Szabo et al., 1995, Curr. Opin. Struct. Biol.5:699-705 and on-line resources provide by BIAcore International AB(Uppsala, Sweden).

Information from SPR can be used to provide an accurate and quantitativemeasure of the equilibrium dissociation constant (K_(D)), and kineticparameters, including K_(on) and K_(off), for the binding of a bindingprotein to a target. Such data can be used to compare differentbiomolecules. For example, selected proteins from an expression librarycan be compared to identify proteins that have high affinity for thetarget or that have a slow K_(off). This information can also be used todevelop structure-activity relationships (SAR). For example, the kineticand equilibrium binding parameters of matured versions of a parentprotein can be compared to the parameters of the parent protein. Variantamino acids at given positions can be identified that correlate withparticular binding parameters, e.g., high affinity and slow K_(off).This information can be combined with structural modeling (e.g., usinghomology modeling, energy minimization, or structure determination byx-ray crystallography or NMR). As a result, an understanding of thephysical interaction between the protein and its target can beformulated and used to guide other design processes.

As a further example, cellular assays may be used. Binding proteins canbe screened for ability to bind to cells which transiently or stablyexpress and display the target of interest on the cell surface. Forexample, Galectin-9 binding proteins can be fluorescently labeled andbinding to Galectin-9 in the presence or absence of antagonisticantibody can be detected by a change in fluorescence intensity usingflow cytometry e.g., a FACS machine.

Antigen-binding fragments of an intact antibody (full-length antibody)can be prepared via routine methods. For example, F(ab′)2 fragments canbe produced by pepsin digestion of an antibody molecule, and Fabfragments that can be generated by reducing the disulfide bridges ofF(ab′)2 fragments.

Genetically engineered antibodies, such as humanized antibodies,chimeric antibodies, single-chain antibodies, and bi-specificantibodies, can be produced via, e.g., conventional recombinanttechnology. In one example, DNA encoding a monoclonal antibodiesspecific to a target antigen can be readily isolated and sequenced usingconventional procedures (e.g., by using oligonucleotide probes that arecapable of binding specifically to genes encoding the heavy and lightchains of the monoclonal antibodies). Once isolated, the DNA may beplaced into one or more expression vectors, which are then transfectedinto host cells such as E. coli cells, simian COS cells, Chinese hamsterovary (CHO) cells, or myeloma cells that do not otherwise produceimmunoglobulin protein, to obtain the synthesis of monoclonal antibodiesin the recombinant host cells. See, e.g., PCT Publication No. WO87/04462. The DNA can then be modified, for example, by substituting thecoding sequence for human heavy and light chain constant domains inplace of the homologous murine sequences, Morrison et al., (1984) Proc.Nat. Acad. Sci. 81:6851, or by covalently joining to the immunoglobulincoding sequence all or part of the coding sequence for anon-immunoglobulin polypeptide. In that manner, genetically engineeredantibodies, such as “chimeric” or “hybrid” antibodies; can be preparedthat have the binding specificity of a target antigen.

Techniques developed for the production of “chimeric antibodies” arewell known in the art. See, e.g., Morrison et al. (1984) Proc. Natl.Acad. Sci. USA 81, 6851; Neuberger et al. (1984) Nature 312, 604; andTakeda et al. (1984) Nature 314:452.

Methods for constructing humanized antibodies are also well known in theart. See, e.g., Queen et al., Proc. Natl. Acad. Sci. USA, 86:10029-10033(1989). In one example, variable regions of V_(H) and V_(L) of a parentnon-human antibody are subjected to three-dimensional molecular modelinganalysis following methods known in the art. Next, framework amino acidresidues predicted to be important for the formation of the correct CDRstructures are identified using the same molecular modeling analysis. Inparallel, human V_(H) and V_(L) chains having amino acid sequences thatare homologous to those of the parent non-human antibody are identifiedfrom any antibody gene database using the parent V_(H) and V_(L)sequences as search queries. Human V_(H) and V_(L) acceptor genes arethen selected.

The CDR regions within the selected human acceptor genes can be replacedwith the CDR regions from the parent non-human antibody or functionalvariants thereof. When necessary, residues within the framework regionsof the parent chain that are predicted to be important in interactingwith the CDR regions (see above description) can be used to substitutefor the corresponding residues in the human acceptor genes.

A single-chain antibody can be prepared via recombinant technology bylinking a nucleotide sequence coding for a heavy chain variable regionand a nucleotide sequence coding for a light chain variable region.Preferably, a flexible linker is incorporated between the two variableregions. Alternatively, techniques described for the production ofsingle chain antibodies (U.S. Pat. Nos. 4,946,778 and 4,704,692) can beadapted to produce a phage or yeast scFv library and scFv clonesspecific to Galectin-9 can be identified from the library followingroutine procedures. Positive clones can be subjected to furtherscreening to identify those that inhibit Galectin-9 activity.

Antibodies obtained following a method known in the art and describedherein can be characterized using methods well known in the art. Forexample, one method is to identify the epitope to which the antigenbinds, or “epitope mapping.” There are many methods known in the art formapping and characterizing the location of epitopes on proteins,including solving the crystal structure of an antibody-antigen complex,competition assays, gene fragment expression assays, and syntheticpeptide-based assays, as described, for example, in Chapter 11 of Harlowand Lane, Using Antibodies, a Laboratory Manual, Cold Spring HarborLaboratory Press, Cold Spring Harbor, N.Y., 1999. In an additionalexample, epitope mapping can be used to determine the sequence, to whichan antibody binds. The epitope can be a linear epitope, i.e., containedin a single stretch of amino acids, or a conformational epitope formedby a three-dimensional interaction of amino acids that may notnecessarily be contained in a single stretch (primary structure linearsequence). Peptides of varying lengths (e.g., at least 4-6 amino acidslong) can be isolated or synthesized (e.g., recombinantly) and used forbinding assays with an antibody. In another example, the epitope towhich the antibody binds can be determined in a systematic screening byusing overlapping peptides derived from the target antigen sequence anddetermining binding by the antibody. According to the gene fragmentexpression assays, the open reading frame encoding the target antigen isfragmented either randomly or by specific genetic constructions and thereactivity of the expressed fragments of the antigen with the antibodyto be tested is determined. The gene fragments may, for example, beproduced by PCR and then transcribed and translated into protein invitro, in the presence of radioactive amino acids. The binding of theantibody to the radioactively labeled antigen fragments is thendetermined by immunoprecipitation and gel electrophoresis. Certainepitopes can also be identified by using large libraries of randompeptide sequences displayed on the surface of phage particles (phagelibraries). Alternatively, a defined library of overlapping peptidefragments can be tested for binding to the test antibody in simplebinding assays. In an additional example, mutagenesis of an antigenbinding domain, domain swapping experiments and alanine scanningmutagenesis can be performed to identify residues required, sufficient,and/or necessary for epitope binding. For example, domain swappingexperiments can be performed using a mutant of a target antigen in whichvarious fragments of the Galectin-9 polypeptide have been replaced(swapped) with sequences from a closely related, but antigenicallydistinct protein (such as another member of the β-galactoside-bindingsoluble lectin family). By assessing binding of the antibody to themutant Galectin-9, the importance of the particular antigen fragment toantibody binding can be assessed.

Alternatively, competition assays can be performed using otherantibodies known to bind to the same antigen to determine whether anantibody binds to the same epitope as the other antibodies. Competitionassays are well known to those of skill in the art.

In some examples, an anti-Galectin-9 antibody is prepared by recombinanttechnology as exemplified below.

Nucleic acids encoding the heavy and light chain of an anti-Galectin-9antibody as described herein can be cloned into one expression vector,each nucleotide sequence being in operable linkage to a suitablepromoter. In one example, each of the nucleotide sequences encoding theheavy chain and light chain is in operable linkage to a distinctpromoter. Alternatively, the nucleotide sequences encoding the heavychain and the light chain can be in operable linkage with a singlepromoter, such that both heavy and light chains are expressed from thesame promoter. When necessary, an internal ribosomal entry site (IRES)can be inserted between the heavy chain and light chain encodingsequences.

In some examples, the nucleotide sequences encoding the two chains ofthe antibody are cloned into two vectors, which can be introduced intothe same or different cells. When the two chains are expressed indifferent cells, each of them can be isolated from the host cellsexpressing such and the isolated heavy chains and light chains can bemixed and incubated under suitable conditions allowing for the formationof the antibody.

Generally, a nucleic acid sequence encoding one or all chains of anantibody can be cloned into a suitable expression vector in operablelinkage with a suitable promoter using methods known in the art. Forexample, the nucleotide sequence and vector can be contacted, undersuitable conditions, with a restriction enzyme to create complementaryends on each molecule that can pair with each other and be joinedtogether with a ligase. Alternatively, synthetic nucleic acid linkerscan be ligated to the termini of a gene. These synthetic linkers containnucleic acid sequences that correspond to a particular restriction sitein the vector. The selection of expression vectors/promoter would dependon the type of host cells for use in producing the antibodies.

A variety of promoters can be used for expression of the antibodiesdescribed herein, including, but not limited to, cytomegalovirus (CMV)intermediate early promoter, a viral LTR such as the Rous sarcoma virusLTR, HIV-LTR, HTLV-1 LTR, the simian virus 40 (SV40) early promoter, E.coli lac UV5 promoter, and the herpes simplex tk virus promoter.

Regulatable promoters can also be used. Such regulatable promotersinclude those using the lac repressor from E. coli as a transcriptionmodulator to regulate transcription from lac operator-bearing mammaliancell promoters [Brown, M. et al., Cell, 49:603-612 (1987)], those usingthe tetracycline repressor (tetR) [Gossen, M., and Bujard, H., Proc.Natl. Acad. Sci. USA 89:5547-5551 (1992); Yao, F. et al., Human GeneTherapy, 9:1939-1950 (1998); Shockelt, P., et al., Proc. Natl. Acad.Sci. USA, 92:6522-6526 (1995)]. Other systems include FK506 dimer, VP16or p65 using astradiol, RU486, diphenol murislerone, or rapamycin.Inducible systems are available from Invitrogen, Clontech and Ariad.

Regulatable promoters that include a repressor with the operon can beused. In one embodiment, the lac repressor from E. coli can function asa transcriptional modulator to regulate transcription from lacoperator-bearing mammalian cell promoters (M. Brown et al., Cell,49:603-612 (1987); Gossen and Bujard (1992); M. Gossen et al., Natl.Acad. Sci. USA, 89:5547-5551 (1992)) combined the tetracycline repressor(tetR) with the transcription activator (VP 16) to create atetR-mammalian cell transcription activator fusion protein, tTa (tetR-VP16), with the tetO-bearing minimal promoter derived from the humancytomegalovirus (hCMV) major immediate-early promoter to create atetR-tet operator system to control gene expression in mammalian cells.In one embodiment, a tetracycline inducible switch is used. Thetetracycline repressor (tetR) alone, rather than the tetR-mammalian celltranscription factor fusion derivatives can function as potenttrans-modulator to regulate gene expression in mammalian cells when thetetracycline operator is properly positioned downstream for the TATAelement of the CMVIE promoter (Yao et al., Human Gene Therapy,10(16):1392-1399 (2003)). One particular advantage of this tetracyclineinducible switch is that it does not require the use of a tetracyclinerepressor-mammalian cells transactivator or repressor fusion protein,which in some instances can be toxic to cells (Gossen et al., Natl.Acad. Sci. USA, 89:5547-5551 (1992); Shockett et al., Proc. Natl. Acad.Sci. USA, 92:6522-6526 (1995)), to achieve its regulatable effects.

Additionally, the vector can contain, for example, some or all of thefollowing: a selectable marker gene, such as the neomycin gene forselection of stable or transient transfectants in mammalian cells;enhancer/promoter sequences from the immediate early gene of human CMVfor high levels of transcription; transcription termination and RNAprocessing signals from SV40 for mRNA stability; SV40 polyoma origins ofreplication and ColE1 for proper episomal replication; internal ribosomebinding sites (IRESes), versatile multiple cloning sites; and T7 and SP6RNA promoters for in vitro transcription of sense and antisense RNA.Suitable vectors and methods for producing vectors containing transgenesare well known and available in the art.

Examples of polyadenylation signals useful to practice the methodsdescribed herein include, but are not limited to, human collagen Ipolyadenylation signal, human collagen II polyadenylation signal, andSV40 polyadenylation signal.

One or more vectors (e.g., expression vectors) comprising nucleic acidsencoding any of the antibodies may be introduced into suitable hostcells for producing the antibodies. The host cells can be cultured undersuitable conditions for expression of the antibody or any polypeptidechain thereof. Such antibodies or polypeptide chains thereof can berecovered by the cultured cells (e.g., from the cells or the culturesupernatant) via a conventional method, e.g., affinity purification. Ifnecessary, polypeptide chains of the antibody can be incubated undersuitable conditions for a suitable period of time allowing forproduction of the antibody.

In some embodiments, methods for preparing an antibody described hereininvolve a recombinant expression vector that encodes both the heavychain and the light chain of an anti-Galectin-9 antibody, as alsodescribed herein. The recombinant expression vector can be introducedinto a suitable host cell (e.g., a dhfr− CHO cell) by a conventionalmethod, e.g., calcium phosphate-mediated transfection. Positivetransformant host cells can be selected and cultured under suitableconditions allowing for the expression of the two polypeptide chainsthat form the antibody, which can be recovered from the cells or fromthe culture medium. When necessary, the two chains recovered from thehost cells can be incubated under suitable conditions allowing for theformation of the antibody.

In one example, two recombinant expression vectors are provided, oneencoding the heavy chain of the anti-Galectin-9 antibody and the otherencoding the light chain of the anti-Galectin-9 antibody. Both of thetwo recombinant expression vectors can be introduced into a suitablehost cell (e.g., dhfr− CHO cell) by a conventional method, e.g., calciumphosphate-mediated transfection. Alternatively, each of the expressionvectors can be introduced into a suitable host cells. Positivetransformants can be selected and cultured under suitable conditionsallowing for the expression of the polypeptide chains of the antibody.When the two expression vectors are introduced into the same host cells,the antibody produced therein can be recovered from the host cells orfrom the culture medium. If necessary, the polypeptide chains can berecovered from the host cells or from the culture medium and thenincubated under suitable conditions allowing for formation of theantibody. When the two expression vectors are introduced into differenthost cells, each of them can be recovered from the corresponding hostcells or from the corresponding culture media. The two polypeptidechains can then be incubated under suitable conditions for formation ofthe antibody.

Standard molecular biology techniques are used to prepare therecombinant expression vector, transfect the host cells, select fortransformants, culture the host cells and recovery of the antibodiesfrom the culture medium. For example, some antibodies can be isolated byaffinity chromatography with a Protein A or Protein G coupled matrix.

Any of the nucleic acids encoding the heavy chain, the light chain, orboth of an anti-Galectin-9 antibody as described herein, vectors (e.g.,expression vectors) containing such; and host cells comprising thevectors are within the scope of the present disclosure.

Anti-Galectin-9 antibodies thus prepared can be can be characterizedusing methods known in the art, whereby reduction, amelioration, orneutralization of Galectin-9 biological activity is detected and/ormeasured. For example, an ELISA-type assay may be suitable forqualitative or quantitative measurement of Galectin-9 inhibition ofDectin-1 or TIM-3 signaling.

The bioactivity of an anti-Galectin-9 antibody can verified byincubating a candidate antibody with Dectin-1 and Galectin-9, andmonitoring any one or more of the following characteristics: (a) bindingbetween Dectin-1 and Galectin-9 and inhibition of the signalingtransduction mediated by the binding; (b) preventing, ameliorating, ortreating any aspect of a solid tumor; (c) blocking or decreasingDectin-1 activation; (d) inhibiting (reducing) synthesis, production orrelease of Galectin-9. Alternatively, TIM-3 can be used to verify thebioactivity of an anti-Galectin-9 antibody using the protocol describedabove. Alternatively, CD206 can be used to verify the bioactivity of ananti-Galectin-9 antibody using the protocol described above.

Additional assays to determine bioactivity of an anti-Galectin-9antibody include measurement of CD8+ and CD4+(conventional) T-cellactivation (in an in vitro or in vivo assay, e.g., by measuringinflammatory cytokine levels, e.g., IFNgamma, TNFalpha, CD44, ICOSgranzymeB, Perforin, IL2 (upregulation); CD26L and IL-10(downregulation)); measurement of reprogramming of macrophages (in vitroor in vivo), e.g., from the M2 to the M1 phenotype (e.g., increasedMHCII, reduced CD206, increased TNF-alpha and iNOS). Alternatively,levels of ADCC can be assessed, e.g., in an in vitro assay, as describedherein.

Methods of Treatment

The present disclosure provides pharmaceutical compositions comprisingat least one anti-Galectin-9 antibody described herein or antigenbinding fragment thereof and uses of such for inhibiting or reducing asignaling mediated by Galectin-9 or eliminating or reducing Galectin-9positive cells. Any of the anti-Galectin-9 antibodies described hereincan be used in any of the methods described herein. In some embodiments,the anti-Galentin-9 antibody is selected from G9.1-1, G9.1-2, G9.1-3,G9.1-4, G9.1-5, G9.1-6, G9.1-7, G9.1-8, G9.1-9, G9.1-10, G9.1-11,G9.1-8m1, G9.1-8m2, G9.1-8m3, G9.1-8m4, G9.1-8m5, G9.1-8m6, G9.1-8m7,G9.1-8m8, G9.1-8m9, G9.1-8m10, G9.1-8m11, G9.1-8m12, G9.1-8m13, andG9.1-8m14, or combinations thereof. In some embodiments, theanti-Galectin-9 antibody is selected from G9.2-1, G9.2-2, G9.2-3,G9.2-4, G9.2-5, G9.2-6, G9.2-7, G9.2-8, G9.2-9, G9.2-10, G9.2-11,G9.2-12, G9.2-13, G9.2-14, G9.2-15, G9.2-16, G9.2-17, G9.2-17mut6,G9.2-18, G9.2-19, G9.2-20, G9.2-21, G9.2-22, G9.2-23, G9.2-24, G9.2-25,G9.2-26, and G9.2-low affinity binder, or combinations thereof.Non-limiting examples of such antibodies include for example antibody9.2-17 or 9.1-8mut13. Such antibodies can be used for treating diseasesassociated with Galectin-9. In some aspects, the invention providesmethods of treating cancer. In some embodiments, the present disclosuremethods for reducing, ameliorating, or eliminating one or moresymptom(s) associated with cancer.

In some embodiments, the disclosure provides a method for treatingcancer in a subject, the method comprising administering to a subject inneed thereof an effective amount of an anti-Galectin-9 antibodydescribed herein or antigen binding fragment thereof. In someembodiments, the disclosure provides a method for treating cancer in asubject, the method comprising administering to a subject in needthereof an effective amount of a pharmaceutical composition comprisingan anti-Galectin-9 antibody described herein or antigen binding fragmentthereof. In some embodiments, the anti-Galectin-9 antibody is selectedfrom the group consisting of G9.1-1, G9.1-2, G9.1-3, G9.1-4, G9.1-5,G9.1-6, G9.1-7, G9.1-8, G9.1-9, G9.1-10, G9.1-11, G9.1-8m1, G9.1-8m2,G9.1-8m3, G9.1-8m4, G9.1-8m5, G9.1-8m6, G9.1-8m7, G9.1- 8m8, G9.1-8m9,G9.1-8m10, G9.1-8m11, G9.1-8m12, G9.1-8m13, and G9.1-8m14 antibodies, orcombinations thereof. In some embodiments, the anti-Galectin-9 antibodyis selected from the group consisting of G9.2-1, G9.2-2, G9.2-3, G9.2-4,G9.2-5, G9.2-6, G9.2-7, G9.2-8, G9.2-9, G9.2-10, G9.2-11, G9.2-12,G9.2-13, G9.2-14, G9.2-15, G9.2-16, G9.2-17, G9.2-17mut6, G9.2-18,G9.2-19, G9.2-20, G9.2-21, G9.2-22, G9.2-23, G9.2-24, G9.2-25, G9.2-26,and G9.2-low affinity binder antibodies, or combinations thereof.Non-limiting examples of such antibodies include for example antibody9.2-17 or 9.1-8mut13.

Given that pro-tumor action of Galectin-9 is mediated throughinteraction with immune cells (i.e., interactions with with lymphoidcells via TIM-3, CD44, and 41BB, and with macrophages via dectin-1 andCD206) and given that Galectin-9 is expressed in a large number oftumors, targeting Galectin-9, e.g., using a Galectin-9 binding antibodyto inhibit interaction with its receptors, provides a therapeuticapproach that can be applied across a variety of different tumor types.

In some embodiments, the cancer is selected from adrenal cancer,adrenocortical carcinoma, anal cancer, appendix cancer, bile ductcancer, bladder cancer, bone cancer (e.g., Ewing sarcoma tumors,osteosarcoma, malignant fibrous histiocytoma), brain cancer (e.g.,astrocytomas, brain stem glioma, craniopharyngioma, ependymoma),bronchial tumors, cholangiocarcinoma, cholangiosarcoma, central nervoussystem tumors, breast cancer, Castleman disease, cervical cancer, coloncancer, rectal cancer, colorectal cancer, endometrial cancer, esophagealcancer, eye cancer, gallbladder cancer, gastrointestinal cancer,gastrointestinal carcinoid tumors, gastrointestinal stromal tumors,genitourinary cancers, gestational trophoblastic disease, heart cancer,Kaposi sarcoma, kidney cancer, laryngeal cancer, hypopharyngeal cancer,leukemia (e.g., acute lymphoblastic leukemia, acute myeloid leukemia,chronic lymphocytic leukemia, chronic myelogenous leukemia), livercancer, lung cancer (for example, non-small cell lung cancer, NSCLC, andsmall cell lung cancer, SCLC), lymphoma (e.g., AIDS-related lymphoma,Burkitt lymphoma, cutaneous T cell lymphoma, Hogkin lymphoma, Non-Hogkinlymphoma, primary central nervous system lymphoma), malignantmesothelioma, multiple myeloma, myelodysplastic syndrome, nasal cavitycancer, paranasal sinus cancer, pancreatic duct adenocarcinoma (PDA)nasopharyngeal cancer, neuroblastoma, oral cavity cancer, oropharyngealcancer, osteosarcoma, ovarian cancer, pancreatic cancer, penile cancer,pituitary tumors, prostate cancer, retinoblastoma, rhabdomyosarcoma,rhabdoid tumor, salivary gland cancer, sarcoma, skin cancer (e.g., basalcell carcinoma, melanoma), squamous cell head and neck cancer, smallintestine cancer, stomach cancer, teratoid tumor, testicular cancer,throat cancer, thymus cancer, thyroid cancer, unusual childhood cancers,upper and lower gastrointestinal malignancies (including, but notlimited to, esophageal, gastric, and hepatobiliary cancer), urethralcancer, uterine cancer, uterine sarcoma, vaginal cancer, vulvar cancer,Waldenstrom macroglobulinemia, and Wilms tumor. In some embodiments, thecancer is selected from hematological malignancies include acutelymphoblastic leukemia, chronic lymphocytic leukemia, lymphomas,multiple myeloma, acute myelogenous leukemia, chronic myelogenousleukemia, myelodysplastic syndromes and the myeloproliferativeneoplasms, such as essential thrombocythemia, polycythemia vera,myelofibrosis, and gallbladder cancer (adenocarcinomas or squamous cellcarcinoma). In some embodiments, the symptom(s) associated thereofinclude, but are not limited to, anemia, loss of appetite, irritation ofbladder lining, bleeding and bruising (thrombocytopenia), changes intaste or smell, constipation, diarrhea, dry mouth, dysphagia, edema,fatigue, hair loss (alopecia), infection, infertility, lymphedema, mouthsores, nausea, pain, peripheral neuropathy, tooth decay, urinary tractinfections, and/or problems with memory and concentration. The methodmay comprise preparing a pharmaceutical composition with ananti-Galectin-9 antibody described herein, and administering thepharmaceutical composition to a subject in a therapeutically effectiveamount.

In some embodiments, the disclosure provides a method for treating gallbladder cancer in a subject, the method comprising administering to asubject in need thereof an effective amount of an anti-Galectin-9antibody described herein, e.g., in Table 1 or Table 2 herein, includingbut not limited to, 9.1-8m13 and/or 9.2-17, or an antigen bindingfragment thereof.

In certain embodiments, administering the pharmaceutical composition,e.g., one or more of the anti-Galectin-9 antibodies described herein,e.g., in Table 1 and/or Table 2, including, but not limited to, 9.2-17and 9.1-8m13, to the subject reduces cell proliferation, tumor growth,and/or tumor volume in a subject, or reduces the number of metastaticlesions over time. In some embodiments, administering the compositionresults in complete response, partial response, or stable disease.

Pancreatic ductal adenocarcinoma (PDA) is a devastating disease with fewlong-term survivors (Yadav et al., Gastroenterology, 2013, 144,1252-1261). Inflammation is paramount in PDA progression as oncogenicmutations alone, in the absence of concomitant inflammation, areinsufficient for tumorigenesis (Guerra et al., Cancer Cell, 2007, 11,291-302). Innate and adaptive immunity cooperate to promote tumorprogression in PDA. In particular, specific innate immune subsets withinthe tumor microenvironment (TME) are apt at educating adaptive immuneeffector cells towards a tumor-permissive phenotype. Antigen presentingcell (APC) populations, including M2-polarized tumor-associatedmacrophages (TAMs) and myeloid dendritic cells (DC), induce thegeneration of immune suppressive Th2 cells in favor of tumor-protectiveTh1 cells (Ochi et al., J of Exp Med., 2012, 209, 1671-1687; Zhu et al.,Cancer Res., 2014, 74, 5057-5069). Similarly, it has been shown thatmyeloid derived suppressor cells (MDSC) negate anti-tumor CD8⁺ cytotoxicT-Lymphocyte (CTL) responses in PDA and promote metastatic progression(Connolly et al., J Leuk Biol., 2010, 87, 713-725; Pylayeva-Gupta etal., Cancer Cell, 2012, 21, 836-847; Bayne et al., Cancer Cell, 2012,21, 822-835).

Recently, dectin-1 on macrophages was shown to bind galectin-9 inpancreatic ductal adenocarcinoma (PDA) (Daley et al., 2017). Removal ofdectin-1 signaling (in Dectin−/− mice) resulted in a decrease in tumorinfiltration of M2 type (suppressive CD206+) macrophages. Additionally,antibody-based Galectin-9 neutralization only enhanced T cell activationin Dectin-1+/+ hosts, indicating that Galectin-9 exerts primaryimmune-suppressive effects specific to Dectin-1 signaling. Uponinterruption of the Dectin-1-Galectin-9 axis, CD4+ and CD8+ Tcells—which are dispensable to PDA progression in hosts with an intactsignaling axis—became reprogrammed into indispensable mediators ofanti-tumor immunity. Without wishing to be bound by theory, blockingGalectin-9-Dectin-1 signaling presents one exemplary mechanism (inaddition to TIM-3 and other signaling pathways) that could underlie astrong anti-tumor response a Galectin-9 targeting immunotherapy approachin PDA e.g., by administering an antibody that binds to Galectin-9, suchas those described herein.

In some embodiments, the disclosure provides a method for treatingpancreatic ductal adenocarcinoma (PDA) in a subject, the methodcomprising administering to a subject in need thereof an effectiveamount of an anti-Galectin-9 antibody described herein or antigenbinding fragment thereof. In some embodiments, the anti-Galentin-9antibody is selected from the group consisting of G9.1-1, G9.1-2,G9.1-3, G9.1-4, G9.1-5, G9.1-6, G9.1-7, G9.1-8, G9.1-9, G9.1-10,G9.1-11, G9.1-8m1, G9.1-8m2, G9.1-8m3, G9.1-8m4, G9.1-8m5, G9.1-8m6,G9.1-8m7, G9.1- 8m8, G9.1-8m9, G9.1-8m10, G9.1-8m11, G9.1-8m12,G9.1-8m13, and G9.1-8m14 antibodies, and combinations thereof. In someembodiments, the anti-Galectin-9 antibody is selected from the groupconsisting of G9.2-1, G9.2-2, G9.2-3, G9.2-4, G9.2-5, G9.2-6, G9.2-7,G9.2-8, G9.2-9, G9.2-10, G9.2-11, G9.2-12, G9.2-13, G9.2-14, G9.2-15,G9.2-16, G9.2-17, G9.2-17mut6, G9.2-18, G9.2-19, G9.2-20, G9.2-21,G9.2-22, G9.2-23, G9.2-24, G9.2-25, G9.2-26, and G9.2-low affinitybinder antibodies, and combinations thereof. Non-limiting examples ofsuch antibodies include for example antibody 9.2-17 or 9.1-8mut13. Inany of these methods of treatment, the anti-Galectin-9 antibody isantibody 9.2-17 and/or antibody 9.1-8mut13.

In some embodiments, the disclosure provides the use of ananti-Galectin-9 antibody as a medicament for the treatment of cancer,wherein the anti-Galectin-9 antibody is any of the antibodies describedherein in Table 1 and/or Table 2, and wherein the cancer is pancreaticductal adenocarcinoma (PDA). In some embodiments, the disclosureprovides the use of an anti-Galectin-9 antibody as a medicament for thetreatment of cancer, wherein the anti-Galectin-9 antibody is antibody9.1-8m13 and/or 9.2-17, and wherein the cancer is pancreatic ductaladenocarcinoma (PDA). Colorectal cancer (CRC), also known as bowelcancer, colon cancer, or rectal cancer, is any cancer affecting thecolon and the rectum. CRC is known to be driven by genetic alterationsof tumor cells and is also influenced by tumor-host interactions. Recentreports have demonstrated a direct correlation between the densities ofcertain T lymphocyte subpopulations and a favorable clinical outcome inCRC, supporting a major role of T-cell-mediated immunity in repressingtumor progression of CRC.

Tim-3, as noted elsewhere herein, is an immune regulatory molecule,which triggers downstream cascade events upon stimulation by galectin-9(Zhu C, et al. The Tim-3 ligand galectin-9 negatively regulates T helpertype 1 immunity; Nature immunology. 2005; 6:1245-1252). Tim-3 has beenfound to be a critical mediator in CRC progression (Yu et al; Tim-3 isupregulated in human colorectal carcinoma and associated with tumorprogression; Mol Med Rep. 2017 February; 15(2): 689-695). In this study,expression of Tim-3 was significantly associated with tumor size(P=0.007), tumor-node-metastasis staging (P<0.0001) and distantmetastasis (P<0.0001). Additionally, increased Tim-3 expression isassociated with M2 macrophage polarization in colon cancer and promotestumor growth. Blockade of the Tim-3 pathway inhibited both thepolarization of tumor-supporting macrophages and colon cancer growth(Jiang et al., Tim-3 promotes tumor-promoting M2 macrophage polarizationby binding to STAT1 and suppressing the STAT1-miR-155 signaling axis;Oncoimmunology, 2016 Aug. 3; 5(9):e1211219). Given these findings andhigh expression of Galectin-9 observed in colorectal cancers (Lahm etal., J. Cancer Res. Clin. Oncol. 2001; 127:375-386), modulating theGalectin-9/TIM-3 axis by inhibiting the interaction between Galectin-9and TIM-3, e.g., by administrating an antibody that binds to Galectin-9,is a novel approach to treating such cancers in the clinic which mayresult in improved outcomes.

In some embodiments, the disclosure provides a method for treatingcolorectal cancer (CRC) in a subject, the method comprisingadministering to a subject in need thereof an effective amount of ananti-Galectin-9 antibody described herein or antigen binding fragmentthereof. In some embodiments, the anti-Galentin-9 antibody is selectedfrom the group consisting of G9.1-1, G9.1-2, G9.1-3, G9.1-4, G9.1-5,G9.1-6, G9.1-7, G9.1-8, G9.1-9, G9.1-10, G9.1-11, G9.1-8m1, G9.1-8m2,G9.1-8m3, G9.1-8m4, G9.1-8m5, G9.1-8m6, G9.1-8m7, G9.1- 8m8, G9.1-8m9,G9.1-8m10, G9.1-8m11, G9.1-8m12, G9.1-8m13, and G9.1-8m14 antibodies,and combinations thereof. In some embodiments, the anti-Galectin-9antibody is selected from the group consisting of G9.2-1, G9.2-2,G9.2-3, G9.2-4, G9.2-5, G9.2-6, G9.2-7, G9.2-8, G9.2-9, G9.2-10,G9.2-11, G9.2-12, G9.2-13, G9.2-14, G9.2-15, G9.2-16, G9.2-17,G9.2-17mut6, G9.2-18, G9.2-19, G9.2-20, G9.2-21, G9.2-22, G9.2-23,G9.2-24, G9.2-25, G9.2-26, and G9.2-low affinity binder antibodies, andcombinations thereof. Non-limiting examples of such antibodies includefor example antibody 9.2-17 or 9.1-8mut13. In any of these methods oftreatment, the anti-Galectin-9 antibody is antibody 9.2-17 and/orantibody 9.1-8mut13.

In some embodiments, the disclosure provides the use of ananti-Galectin-9 antibody as a medicament for the treatment of cancer,wherein the anti-Galectin-9 antibody is any of the antibodies describedherein in Table 1 and/or Table 2, and wherein the cancer is colorectalcancer (CRC). In some embodiments, the disclosure provides the use of ananti-Galectin-9 antibody as a medicament for the treatment of cancer,wherein the anti-Galectin-9 antibody is antibody 9.1-8m13 and/or 9.2-17,and wherein the cancer is colorectal cancer (CRC).

Melanoma is the deadliest form of skin cancer and has been increasing inincidence for the past 30 years, especially in young adults. Recentadvances have resulted in the development of numerous immune-activatingtherapies that have greatly improved patient survival.

Accumulation of genetic disorders, most frequently mutations in B-Rafand N-Ras, in the melanocyte are a hallmark of melanoma(Rodriguez-Cerdeira et al., Advances in Immunotherapy for Melanoma: AComprehensive Review; Mediators Inflamm. 2017; 2017: 3264217, andreferences therein). However, the interaction between themicroenvironment is necessary for these alterations to result in thetransformation of a dysplastic melanocyte into a melanoma cell. Themicroenvironment then also further promotes invasion and metastasis. Newtherapeutic strategies including CTLA-4, PD-1 and PD-L1/2 blockers, havebeen developed and have dramatically improved outcomes for melanomapatients (Farkona et al., Cancer immunotherapy: the beginning of the endof cancer? BMC Med. 2016; 14:73). However, these therapies depend on thepresence of a functional immune system, which is suppressed in patientswith advanced cancer, and new methods to reactivate this suppressedsystemic immunity are needed to further improve outcomes for melanomapatients.

In patients with metastatic melanoma, high blood levels of galectin-9are correlated with worse overall survival and a bias towards a Th2inflammatory state supportive of tumor growth. Additionally, galectin-9is co-localized with the M2 macrophage population in metastatic melanomaand soluble forms of galectin-9 in the blood correspond with poorsurvival (Enninga et al., Melanoma Res. 2016 October; 26(5):429-41).Association of Galectin-9 with M2 macrophages was found to be due toGalectin-9 ligation to CD206 on M2 macrophages, which resulted inpro-tumor phenotype in the local microenvironment. Accordingly, bothGalectin-9/dectin-1 and Galectin-9/CD206 interactions may promotemacrophage mediated immune suppressive effects. Without wishing to bebound by theory, these findings indicate that inhibitingGalectin-9/dectin-1 and Galectin-9/CD206 interactions, e.g., byadministering an antibody that binds to Galectin-9, may present arationale for employing anti-Galectin-9 antibodies in a therapeuticapproach in melanoma, which will lead to improved overall survival, inpatients, including but not limited to those refractory to anti-CTLA-4,PD-1 and PD-L1/2 therapies.

In some embodiments, the disclosure provides a method for treatingmelanoma in a subject, the method comprising administering to a subjectin need thereof an effective amount of an anti-Galectin-9 antibodydescribed herein or antigen binding fragment thereof. In someembodiments, the anti-Galentin-9 antibody is selected from the groupconsisting of G9.1-1, G9.1-2, G9.1-3, G9.1-4, G9.1-5, G9.1-6, G9.1-7,G9.1-8, G9.1-9, G9.1-10, G9.1-11, G9.1-8m1, G9.1-8m2, G9.1-8m3,G9.1-8m4, G9.1-8m5, G9.1-8m6, G9.1-8m7, G9.1-8m8, G9.1-8m9, G9.1-8m10,G9.1-8m11, G9.1-8m12, G9.1-8m13, and G9.1-8m14 antibodies, andcombinations thereof. In some embodiments, the anti-Galectin-9 antibodyis selected from the group consisting of G9.2-1, G9.2-2, G9.2-3, G9.2-4,G9.2-5, G9.2-6, G9.2-7, G9.2-8, G9.2-9, G9.2-10, G9.2-11, G9.2-12,G9.2-13, G9.2-14, G9.2-15, G9.2-16, G9.2-17, G9.2-17mut6, G9.2-18,G9.2-19, G9.2-20, G9.2-21, G9.2-22, G9.2-23, G9.2-24, G9.2-25, G9.2-26,and G9.2-low affinity binder antibodies, and combinations thereof.Non-limiting examples of such antibodies include for example antibody9.2-17 or 9.1-8mut13. In any of these methods of treatment, theanti-Galectin-9 antibody is antibody 9.2-17 and/or antibody 9.1-8mut13.

In some embodiments, the disclosure provides the use of ananti-Galectin-9 antibody as a medicament for the treatment of cancer,wherein the anti-Galectin-9 antibody is any of the antibodies describedherein in Table 1 and/or Table 2, and wherein the cancer is melanoma. Insome embodiments, the disclosure provides the use of an anti-Galectin-9antibody as a medicament for the treatment of cancer, wherein theanti-Galectin-9 antibody is antibody 9.1-8m13 and/or 9.2-17, and whereinthe cancer is melanoma.

Cholangiocarcinoma (CCA) is an epithelial cancer that forms in the bileducts and is the most common biliary malignancy and the second mostcommon hepatic malignancy after hepatocellular carcinoma and the overallincidence of cholangiocarcinoma has increased progressively worldwideover the past four decades. CCAs are classified into three subtypesbased on their anatomic location, intrahepatic cholangiocarcinoma(iCCA), perihilar CCA (pCCA), and distal CCA (dCCA) (see, e.g.,Loeuillard et al., Animal models of cholangiocarcinoma; Biochim BiophysActa Mol Basis Dis. 2018 Apr. 5, and Rizvi et al.,Cholangiocarcinoma—evolving concepts and therapeutic strategies; Nat RevClin Oncol. 2018 February; 15(2): 95-111).

In a restrospective immune profiling study in of 99 surgically resectediHCC, TIM-3-positive staining of centrally located, tumor infiltrativelymphocytes was observed, at levels 3 times greater than PD-1 staining.Overall survival was significantly associated with lower numbers ofTIM-3 tumor infiltrating lymphocytes(ascopubs.org/doi/abs/10.1200/JC0.2018.36.15_suppl.12049). Accordingly,reducing TIM-3 activity or signaling, e.g., by inhibiting theGal-9/Tim-3 interaction in an immunotherapeutic approach, e.g., byadministering an anti-Galectin-9 antibody such as one or more of theanti-Galectin-9 antibodies described herein, may have a positive impacton overall survival.

In some embodiments, the disclosure provides a method for treatingcholangiocarcinoma in a subject, the method comprising administering toa subject in need thereof an effective amount of an anti-Galectin-9antibody described herein or antigen binding fragment thereof. In someembodiments, the anti-Galentin-9 antibody is selected from the groupconsisting of G9.1-1, G9.1-2, G9.1-3, G9.1-4, G9.1-5, G9.1-6, G9.1-7,G9.1-8, G9.1-9, G9.1-10, G9.1-11, G9.1-8m1, G9.1-8m2, G9.1-8m3,G9.1-8m4, G9.1-8m5, G9.1-8m6, G9.1-8m7, G9.1-8m8, G9.1-8m9, G9.1-8m10,G9.1-8m11, G9.1-8m12, G9.1-8m13, and G9.1-8m14 antibodies, andcombinations thereof. In some embodiments, the anti-Galectin-9 antibodyis selected from the group consisting of G9.2-1, G9.2-2, G9.2-3, G9.2-4,G9.2-5, G9.2-6, G9.2-7, G9.2-8, G9.2-9, G9.2-10, G9.2-11, G9.2-12,G9.2-13, G9.2-14, G9.2-15, G9.2-16, G9.2-17, G9.2-17mut6, G9.2-18,G9.2-19, G9.2-20, G9.2-21, G9.2-22, G9.2-23, G9.2-24, G9.2-25, G9.2-26,and G9.2-low affinity binder antibodies, and combinations thereof.Non-limiting examples of such antibodies include for example antibody9.2-17 or 9.1-8mut13. In any of these methods of treatment, theanti-Galectin-9 antibody is antibody 9.2-17 and/or antibody 9.1-8mut13.

In some embodiments, the disclosure provides the use of ananti-Galectin-9 antibody as a medicament for the treatment of cancer,wherein the anti-Galectin-9 antibody is any of the antibodies describedherein in Table 1 and/or Table 2, and wherein the cancer ischolangiocarcinoma. In some embodiments, the disclosure provides the useof an anti-Galectin-9 antibody as a medicament for the treatment ofcancer, wherein the anti-Galectin-9 antibody is antibody 9.1-8m13 and/or9.2-17, and wherein the cancer is cholangiocarcinoma.

Renal Cell Carcinoma (RCC) has the highest mortality rate of thegenitourinary cancers and the incidence of RCC has risen steadily, whilethe outcome remains poor. Approximately 273,000 new cases of kidneycancer are diagnosed worldwide each year. About one third of patientswith localized disease will suffer recurrence or metastasis. Oncemetastasis occurs, malignancy metastasize, the 5-year survival forpatients is less than 10%. Clear-cell renal cell carcinoma (ccRCC) isthe major histological subtype, which accounts for 80-90% of all theRCCs. RCC is sensitive to immunotherapy and targeted therapy whilehighly resistant to both chemotherapy and radiation therapy.

In RCC patients, Gal-9 is expressed at much higher levels in cancerouslesions than the surrounding normal tissue, and patients with highGalectin-9 expression showed more advanced progression of the diseasewith larger tumor size and necrosis (Kawashima et al.; BJU Int. 2014;113:320-332). Gal-9 in tumor tissue of ccRCC patients was significantlypositively associated with tumor size, Fuhrman grade, necrosis, andimpaired clinical outcome including poor survival and early recurrence(Fu et al., Galectin-9 predicts postoperative recurrence and survival ofpatients with clear-cell renal cell carcinoma; Tumour Biol. 2015 August;36(8):5791-9). TIM-3 is also associated with poor prognosis in RCC, andknockdown of TIM-3 suppresses the proliferation and invasion capacity ofccRCC cell lines (Yuan et al., Prognostic implication of Tim-3 in clearcell renal cell carcinoma. Neoplasma. 2014; 61:35-40). Accordingly, theGal-9/TIM-3 axis might play an important role in the development ofrenal cell carcinoma and administration of immunotherapeutic agentswhich inhibit Gal-9 binding to TIM-3, such as the anti-Galectin-9antibodies described herein, e.g., in Table 1 and/or Table 2, may resultin increased survival and lower reoccurrence in RCC.

In some embodiments, the disclosure provides a method for treating renalcancer in a subject, the method comprising administering to a subject inneed thereof an effective amount of an anti-Galectin-9 antibodydescribed herein or antigen binding fragment thereof. In someembodiments, the anti-Galentin-9 antibody is selected from the groupconsisting of G9.1-1, G9.1-2, G9.1-3, G9.1-4, G9.1-5, G9.1-6, G9.1-7,G9.1-8, G9.1-9, G9.1-10, G9.1-11, G9.1-8m1, G9.1-8m2, G9.1-8m3,G9.1-8m4, G9.1-8m5, G9.1-8m6, G9.1-8m7, G9.1-8m8, G9.1-8m9, G9.1-8m10,G9.1-8m11, G9.1-8m12, G9.1-8m13, and G9.1-8m14 antibodies, andcombinations thereof. In some embodiments, the anti-Galectin-9 antibodyis selected from the group consisting of G9.2-1, G9.2-2, G9.2-3, G9.2-4,G9.2-5, G9.2-6, G9.2-7, G9.2-8, G9.2-9, G9.2-10, G9.2-11, G9.2-12,G9.2-13, G9.2-14, G9.2-15, G9.2-16, G9.2-17, G9.2-17mut6, G9.2-18,G9.2-19, G9.2-20, G9.2-21, G9.2-22, G9.2-23, G9.2-24, G9.2-25, G9.2-26,and G9.2-low affinity binder antibodies, and combinations thereof.Non-limiting examples of such antibodies include for example antibody9.2-17 or antibody 9.1-8mut13. In any of these methods of treatment, theanti-Galectin-9 antibody is antibody 9.2-17 and/or antibody 9.1-8mut13.

In some embodiments, the disclosure provides the use of ananti-Galectin-9 antibody as a medicament for the treatment of cancer,wherein the anti-Galectin-9 antibody is any of the antibodies describedherein in Table 1 and/or Table 2, and wherein the cancer is renal cellcarcinoma (RCC). In some embodiments, the disclosure provides the use ofan anti-Galectin-9 antibody as a medicament for the treatment of cancer,wherein the anti-Galectin-9 antibody is antibody 9.1-8m13 and/or 9.2-17,and wherein the cancer is renal cell carcinoma (RCC).

Hepatocellular carcinoma (HCC) is the most common type of primary livercancer. Hepatocellular carcinoma occurs most often in people withchronic liver diseases, such as cirrhosis caused by hepatitis B orhepatitis C infection. HCC is usually accompanied by cirrhotic liverwith extensive lymphocyte infiltration due to chronic viral infection.Many studies have demonstrated that tumor-infiltrating effector CD8+ Tcells and T helper 17 (Th17) cells correlate with improved survivalafter surgical resection of tumors. However, tumor-infiltrating effectorT cells fail to control tumor growth and metastasis Pang et al., Theimmunosuppressive tumor microenvironment in hepatocellular carcinoma;Cancer Immunol Immunother 2009; 58:877-886).

The TIM-3/galectin-9 interaction contributes to immune dysfunction inhuman HCC (Li, et al., Tim-3/galectin-9 signaling pathway mediatesT-cell dysfunction and predicts poor prognosis in patients withhepatitis B virus-associated hepatocellular carcinoma; Hepatology. 2012October; 56(4):1342-51). High Galectin-9 expression is found on myeloidAPCs and high numbers of Tim-3+ T cells are found in HBV-associated HCC,and blocking Tim-3/galectin-9 signaling using TIM-3 antibodies recoverseffector T-cell function in T cells isolated from human HCC. Thus, thetargeting Tim-3/Galectin-9 axis, e.g., by administering anti-Galectin-9antibodies, e.g., such as anti-Galectin-9 antibodies shown in Table 1and Table 2 herein, including, but not limited to, antibody 9.1-8mut13and/or antibody 9.2-17, constitutes a novel immune therapeutic strategyfor treating patients with HBV-associated HCC.

In some embodiments, the disclosure provides a method for treatinghepatocellular carcinoma in a subject, the method comprisingadministering to a subject in need thereof an effective amount of ananti-Galectin-9 antibody described herein or antigen binding fragmentthereof. In some embodiments, the anti-Galentin-9 antibody is selectedfrom the group consisting of G9.1-1, G9.1-2, G9.1-3, G9.1-4, G9.1-5,G9.1-6, G9.1-7, G9.1-8, G9.1-9, G9.1-10, G9.1-11, G9.1-8m1, G9.1-8m2,G9.1-8m3, G9.1-8m4, G9.1-8m5, G9.1-8m6, G9.1-8m7, G9.1- 8m8, G9.1-8m9,G9.1-8m10, G9.1-8m11, G9.1-8m12, G9.1-8m13, and G9.1-8m14 antibodies,and combinations thereof. In some embodiments, the anti-Galectin-9antibody is selected from the group consisting of G9.2-1, G9.2-2,G9.2-3, G9.2-4, G9.2-5, G9.2-6, G9.2-7, G9.2-8, G9.2-9, G9.2-10,G9.2-11, G9.2-12, G9.2-13, G9.2-14, G9.2-15, G9.2-16, G9.2-17,G9.2-17mut6, G9.2-18, G9.2-19, G9.2-20, G9.2-21, G9.2-22, G9.2-23,G9.2-24, G9.2-25, G9.2-26, and G9.2-low affinity binder antibodies, andcombinations thereof. Non-limiting examples of such antibodies includefor example antibody 9.2-17 or 9.1-8mut13. In any of these methods oftreatment, the anti-Galectin-9 antibody is antibody 9.2-17 and/orantibody 9.1-8mut13. Acute myeloid leukemia (AML) is the most commonform of acute leukemia, with an incidence that increases with advancedage. Commonly of unknown etiology, AML can also occur as a result ofexposure to genotoxic agents or following a previous hematologicdisorder. AML is complex, with genetic, epigenetic, and phenotypicheterogeneity (Lowenberg and Rowe, Introduction to the review series onadvances in acute myeloid leukemia (AML); Blood 2016 127:1).

In some embodiments, the disclosure provides the use of ananti-Galectin-9 antibody as a medicament for the treatment of cancer,wherein the anti-Galectin-9 antibody is any of the antibodies describedherein in Table 1 and/or Table 2, and wherein the cancer ishepatocellular carcinoma (HCC). In some embodiments, the disclosureprovides the use of an anti-Galectin-9 antibody as a medicament for thetreatment of cancer, wherein the anti-Galectin-9 antibody is antibody9.1-8m13 and/or 9.2-17, and wherein the cancer is hepatocellularcarcinoma (HCC).

Recent studies suggest that the TIM-3/Gal-9 axis that TIM-3 and Gal-9are connected to the establishment of AML. Malignant stem cells achievedominant clonal selection through acquisition of multiple geneticabnormalities. These genetic abnormalities progressively accumulate inself-renewing hematopoietic stem cells (HSCs), and, as a consequence,these genetically impaired preleukemic HSCs transform into leukemic stemcells (LSCs). As part of this process, preleukemic HSCs outgrow normalHSCs, and finally self-renew at a hematopoietic progenitor cell stage tobecome myeloid LSCs (Walter et al., Clonal architecture of secondaryacute myeloid leukemia; N. Engl. J. Med., 366 (2012), pp. 1090-1098).Kikshige et al., (Kikushige et al., A TIM-3/Gal-9 Autocrine StimulatoryLoop Drives Self-Renewal of Human Myeloid Leukemia Stem Cells andLeukemic Progression (Cell Stem Cell 17; 3(2015), 341-352) observed thatserum Galectin-9 levels were significantly elevated in AML patients andthat the Tim3/Gal-9 axis stimulates an autocrine loop which functions toallow clonal dominancy and self-renewal of LSCs. Gal-9-mediated TIM-3stimulation lead to the induction of LSC self renewal pathways. Of note,since significant upregulation of TIM-3 in HSC and HPC populations, aswell as elevation of serum Gal-9, was observed in patients withpreleukemic myeloid disorders, acquisition of Galectin-9 secretionlikely occurs early during leukemia progression. Accordingly, targetingthe Gal-9/TIM-3 axis, e.g., through the administration of ananti-Galectin-9 antibody, such as one or more of the anti-Galectin-9antibodies described herein, e.g., in Table 1 and/or Table 2, includingantibody 9.1-8mut13 and/or antibody 9.2-17, may constitute a novelapproach to cancer stem cell therapy common to human myeloidmalignancies, and moreover, such therapies may be useful not only toeradicate LSCs in AMLs, but also to prevent progression of preleukemicdisorders into overt AML. Such preleukemic disorders include therefractory cytopenia with multilineage displasia (RCMD) stage inmyelodysplastic syndromes (MDS) or the chronic phase ofmyeloproliferative neoplasms (MPN), including chronic myelogenousleukemia.

In some embodiments, the disclosure provides a method for treating ahematological malignancy in a subject, the method comprisingadministering to a subject in need thereof an effective amount of ananti-Galectin-9 antibody described herein or antigen binding fragmentthereof. In some embodiments, the disclosure provides a method fortreating acute lymphoblastic leukemia in a subject, the methodcomprising administering to a subject in need thereof an effectiveamount of an anti-Galectin-9 antibody described herein or antigenbinding fragment thereof. In some embodiments, the disclosure provides amethod for treating acute myeloid leukemia in a subject, the methodcomprising administering to a subject in need thereof an effectiveamount of an anti-Galectin-9 antibody described herein or antigenbinding fragment thereof.

In some embodiments, the disclosure provides a method for preventingprogression of preleukemic disorders into acute myeloid leukemia in asubject, the method comprising administering to a subject in needthereof an effective amount of an anti-Galectin-9 antibody describedherein or antigen binding fragment thereof. In some embodiments, thepreleukemic disorders comprise RCMD stage in MDS or the chronic phase ofMPN, including chronic myelogenous leukemia. In some embodiments, theanti-Galentin-9 antibody is selected from the group consisting ofG9.1-1, G9.1-2, G9.1-3, G9.1-4, G9.1-5, G9.1-6, G9.1-7, G9.1-8, G9.1-9,G9.1-10, G9.1-11, G9.1-8m1, G9.1-8m2, G9.1-8m3, G9.1-8m4, G9.1-8m5,G9.1-8m6, G9.1-8m7, G9.1-8m8, G9.1-8m9, G9.1-8m10, G9.1-8m11, G9.1-8m12,G9.1-8m13, and G9.1-8m14 antibodies, and combinations thereof. In someembodiments, the anti-Galectin-9 antibody is selected from the groupconsisting of G9.2-1, G9.2-2, G9.2-3, G9.2-4, G9.2-5, G9.2-6, G9.2-7,G9.2-8, G9.2-9, G9.2-10, G9.2-11, G9.2-12, G9.2-13, G9.2-14, G9.2-15,G9.2-16, G9.2-17, G9.2-17mut6, G9.2-18, G9.2-19, G9.2-20, G9.2-21,G9.2-22, G9.2-23, G9.2-24, G9.2-25, G9.2-26, and G9.2-low affinitybinder antibodies, and combinations thereof. Non-limiting examples ofsuch antibodies include for example antibody 9.2-17 or 9.1-8mut13. Inany of these methods of treatment, the anti-Galectin-9 antibody isantibody 9.2-17 and/or antibody 9.1-8mut13.

In some embodiments, the disclosure provides the use of ananti-Galectin-9 antibody as a medicament for the treatment of cancer,wherein the anti-Galectin-9 antibody is any of the antibodies describedherein in Table 1 and/or Table 2, and wherein the cancer is ahematological malignancy. In some embodiments, the disclosure providesthe use of an anti-Galectin-9 antibody as a medicament for the treatmentof cancer, wherein the anti-Galectin-9 antibody is antibody 9.1-8m13and/or 9.2-17, and wherein the cancer is hematological malignancy.

In some embodiments, the disclosure provides the use of ananti-Galectin-9 antibody as a medicament for the treatment of cancer,wherein the anti-Galectin-9 antibody is any of the antibodies describedherein in Table 1 and/or Table 2, and wherein the cancer is AML. In someembodiments, the disclosure provides the use of an anti-Galectin-9antibody as a medicament for the treatment of cancer, wherein theanti-Galectin-9 antibody is antibody 9.1-8m13 and/or 9.2-17, and whereinthe cancer is AML.

In some embodiments, the disclosure provides the use of ananti-Galectin-9 antibody as a medicament for the treatment of cancer,wherein the anti-Galectin-9 antibody is any of the antibodies describedherein in Table 1 and/or Table 2, and wherein the cancer is ALL. In someembodiments, the disclosure provides the use of an anti-Galectin-9antibody as a medicament for the treatment of cancer, wherein theanti-Galectin-9 antibody is antibody 9.1-8m13 and/or 9.2-17, and whereinthe cancer is ALL. In any of the above-described methods, the treatmentmethod further comprises administering to the subject an inhibitor of acheckpoint molecule, an activator of a co-stimulatory receptor, and/oran inhibitor of an innate immune cell target. In some embodiments, thetreatment method further comprises administering to the subject aninhibitor of a checkpoint molecule. In some embodiments, the checkpointmolecule is selected from the group consisting of PD-1, PD-L1, PD-L2,CTLA-4, LAG3, TIM-3 and A2aR. In some embodiments, the treatment methodfurther comprises administering to the subject an inhibitor of anactivator of a co-stimulatory receptor, and/or an inhibitor of an innateimmune cell target. In some embodiments, the co-stimulatory receptor isselected from the group consisting of OX40, GITR, CD137, CD40, CD27, andICOS. In some embodiments, the treatment method further comprisesadministering to the subject an inhibitor of an innate immune celltarget. In some embodiments, the innate immune cell target is selectedfrom the group consisting of KIR, NKG2A, CD96, TLR, and IDO. In someembodiments, the anti-Galectin-9 antibody is selected from the groupconsisting of G9.1-1, G9.1-2, G9.1-3, G9.1-4, G9.1-5, G9.1-6, G9.1-7,G9.1-8, G9.1-9, G9.1-10, G9.1-11, G9.1-8m1, G9.1-8m2, G9.1-8m3,G9.1-8m4, G9.1-8m5, G9.1-8m6, G9.1-8m7, G9.1-8m8, G9.1-8m9, G9.1-8m10,G9.1-8m11, G9.1-8m12, G9.1-8m13, and G9.1-8m14 antibodies, andcombinations thereof. In some embodiments, the anti-Galectin-9 antibodyis selected from the group consisting of G9.2-1, G9.2-2, G9.2-3, G9.2-4,G9.2-5, G9.2-6, G9.2-7, G9.2-8, G9.2-9, G9.2-10, G9.2-11, G9.2-12,G9.2-13, G9.2-14, G9.2-15, G9.2-16, G9.2-17, G9.2-17mut6, G9.2-18,G9.2-19, G9.2-20, G9.2-21, G9.2-22, G9.2-23, G9.2-24, G9.2-25, G9.2-26,and G9.2-low affinity binder antibodies, and combinations thereof.Non-limiting examples of such antibodies include for example antibody9.2-17 or 9.1-8mut13. In any of these methods of treatment, theanti-Galectin-9 antibody is antibody 9.2-17 and/or antibody 9.1-8mut13.In some embodiments, the cancer is selected from pancreatic cancer,e.g., pancreatic ductal adenocarcinoma, cholangiocarcinoma,hepatocellular carcinoma, colorectal cancer, melanoma, renal cellcarcinoma, and acute myeloid leukemia.

In some embodiments, the disclosure provides the use of ananti-Galectin-9 antibody as a medicament for the treatment of cancer incombination with a checkpoint inhibitor molecule, e.g., wherein thecheckpoint inhibitor molecule is selected from the group consisting ofPD-1, PD-L1, PD-L2, CTLA-4, LAG3, TIM-3 and A2aR, wherein theanti-Galectin-9 antibody is any of the antibodies described herein inTable 1 and/or Table 2. In some embodiments, the disclosure provides theuse of an anti-Galectin-9 antibody as a medicament for the treatment ofcancer in combination with a checkpoint molecule, wherein the checkpointinhibitor molecule is selected from the group consisting of PD-1, PD-L1,PD-L2, CTLA-4, LAG3, TIM-3 and A2aR, and wherein the anti-Galectin-9antibody is antibody 9.1-8m13 and/or 9.2-17.

In some embodiments, the disclosure provides the use of ananti-Galectin-9 antibody as a medicament for the treatment of cancer incombination with a co-stimulatory molecule, e.g., wherein theco-stimulatory molecule is selected from the group consisting of OX40,GITR, CD137, CD40, CD27, and ICOS, and wherein the anti-Galectin-9antibody is any of the antibodies described herein in Table 1 and/orTable 2. In some embodiments, the disclosure provides the use of ananti-Galectin-9 antibody as a medicament for the treatment of cancer incombination with a co-stimulatory, wherein the co-stimulatory moleculeis selected from the group consisting of OX40, GITR, CD137, CD40, CD27,and ICOS, wherein the anti-Galectin-9 antibody is antibody 9.1-8m13and/or 9.2-17. In some embodiments, the methods of the presentdisclosure may increase anti-tumor activity (e.g., reduce cellproliferation, tumor growth, tumor volume, and/or tumor burden or loador reduce the number of metastatic lesions over time) by at least about10%, 20%, 25%, 30%, 40%, 50%, 60%, 70%, 75%, 80%, 85%, 90%, 95%, or moreas compared to levels prior to treatment or in a control subject. Insome embodiments, reduction is measured by comparing cell proliferation,tumor growth, and/or tumor volume in a subject before and afteradministration of the pharmaceutical composition. In some embodiments,the method of treating or ameliorating a cancer in a subject allows oneor more symptoms of the cancer to improve by at least about 10%, 20%,30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, or more. Before, during, andafter the administration of the pharmaceutical composition, cancerouscells and/or biomarkers in a subject may be measured in a biologicalsample, such as blood, serum, plasma, urine, peritoneal fluid, and/or abiopsy from a tissue or organ. In some embodiments, the methods mayinclude administration of the compositions of the invention to reducetumor volume, size, load or burden in a subject to an undetectable size,or to less than about 1%, 2%, 5%, 10%, 20%, 25%, 30%, 40%, 50%, 60%,70%, 75%, 80%, or 90% of the subject's tumor volume, size, load orburden prior to treatment. In other embodiments, the methods may includeadministration of the compositions of the invention to reduce the cellproliferation rate or tumor growth rate in a subject to an undetectablerate, or to less than about 1%, 2%, 5%, 10%, 20%, 25%, 30%, 40%, 50%,60%, 70%, 75%, 80%, or 90% of the rate prior to treatment. In otherembodiments, the methods may include administration of the compositionsof the invention to reduce the development of or the number or size ofmetastatic lesions in a subject to an undetectable rate, or to less thanabout 1%, 2%, 5%, 10%, 20%, 25%, 30%, 40%, 50%, 60%, 70%, 75%, 80%, or90% of the rate prior to treatment.

In some embodiments, compositions are provided comprising one or moreanti-Galectin-9 antibodies, which may be used to treat, manage,ameliorate, and/or prevent cancer. In some embodiments, compositions ofthe disclosure comprise two or more anti-Galectin-9 antibodies, alone orin combination with prophylactic agents, therapeutic agents (e.g.,chemotherapy or immunotherapy), and/or pharmaceutically acceptablecarriers and the use thereof are provided. In some embodiments, the oneor more antibodies bind to CRD1. In some embodiments, the one or moreantibodies bind to CRD2. In some embodiments, the one or more antibodiesbind to CRD1. In some embodiments, the compositions can comprise acombinattion of antibodies, some of which bind to CRD1, and some ofwhich bind to CRD2. A non-limiting example of a combination is acombination comprising 9.2-17 and 9.1-8mut1. Antibodies can be combinedin equimolar or non-equimolar amounts.

Pharmaceutical Compositions

The anti-Galectin-9 antibodies, as well as the encoding nucleic acids ornucleic acid sets, vectors comprising such, or host cells comprising thevectors, as described herein can be mixed with a pharmaceuticallyacceptable carrier (excipient) to form a pharmaceutical composition foruse in treating a target disease. “Acceptable” means that the carriermust be compatible with the active ingredient of the composition (andpreferably, capable of stabilizing the active ingredient) and notdeleterious to the subject to be treated. Pharmaceutically acceptableexcipients (carriers) including buffers, which are well known in theart. See, e.g., Remington: The Science and Practice of Pharmacy 20th Ed.(2000) Lippincott Williams and Wilkins, Ed. K. E. Hoover.

The pharmaceutical compositions to be used in the present methods cancomprise pharmaceutically acceptable carriers, excipients, orstabilizers in the form of lyophilized formulations or aqueoussolutions. (Remington: The Science and Practice of Pharmacy 20th Ed.(2000) Lippincott Williams and Wilkins, Ed. K. E. Hoover). Acceptablecarriers, excipients, or stabilizers are nontoxic to recipients at thedosages and concentrations used, and may comprise buffers such asphosphate, citrate, and other organic acids; antioxidants includingascorbic acid and methionine; preservatives (such asoctadecyldimethylbenzyl ammonium chloride; hexamethonium chloride;benzalkonium chloride, benzethonium chloride; phenol, butyl or benzylalcohol; alkyl parabens such as methyl or propyl paraben; catechol;resorcinol; cyclohexanol; 3-pentanol; and m-cresol); low molecularweight (less than about 10 residues) polypeptides; proteins, such asserum albumin, gelatin, or immunoglobulins; hydrophilic polymers such aspolyvinylpyrrolidone; amino acids such as glycine, glutamine,asparagine, histidine, arginine, or lysine; monosaccharides,disaccharides, and other carbohydrates including glucose, mannose, ordextrans; chelating agents such as EDTA; sugars such as sucrose,mannitol, trehalose or sorbitol; salt-forming counter-ions such assodium; metal complexes (e.g. Zn-protein complexes); and/or non-ionicsurfactants such as TWEEN′, PLURONICS' or polyethylene glycol (PEG). Insome examples, the pharmaceutical composition described herein comprisesliposomes containing the antibodies (or the encoding nucleic acids)which can be prepared by methods known in the art, such as described inEpstein, et al., Proc. Natl. Acad. Sci. USA 82:3688 (1985); Hwang, etal., Proc. Natl. Acad. Sci. USA 77:4030 (1980); and U.S. Pat. Nos.4,485,045 and 4,544,545. Liposomes with enhanced circulation time aredisclosed in U.S. Pat. No. 5,013,556. Particularly useful liposomes canbe generated by the reverse phase evaporation method with a lipidcomposition comprising phosphatidylcholine, cholesterol andPEG-derivatized phosphatidylethanolamine (PEG-PE). Liposomes areextruded through filters of defined pore size to yield liposomes withthe desired diameter.

The anti-Galectin-9 antibodies, or the encoding nucleic acid(s), mayalso be entrapped in microcapsules prepared, for example, bycoacervation techniques or by interfacial polymerization, for example,hydroxymethylcellulose or gelatin-microcapsules andpoly-(methylmethacylate) microcapsules, respectively, in colloidal drugdelivery systems (for example, liposomes, albumin microspheres,microemulsions, nano-particles and nanocapsules) or in macroemulsions.Such techniques are known in the art, see, e.g., Remington, The Scienceand Practice of Pharmacy 20th Ed. Mack Publishing (2000).

In other examples, the pharmaceutical composition described herein canbe formulated in sustained-release format. Suitable examples ofsustained-release preparations include semipermeable matrices of solidhydrophobic polymers containing the antibody, which matrices are in theform of shaped articles, e.g. films, or microcapsules. Examples ofsustained-release matrices include polyesters, hydrogels (for example,poly(2-hydroxyethyl-methacrylate), or poly(vinyl alcohol)), polylactides(U.S. Pat. No. 3,773,919), copolymers of L-glutamic acid and 7ethyl-L-glutamate, non-degradable ethylene-vinyl acetate, degradablelactic acid-glycolic acid copolymers such as the LUPRON DEPOT′(injectable microspheres composed of lactic acid-glycolic acid copolymerand leuprolide acetate), sucrose acetate isobutyrate, andpoly-D-(−)-3-hydroxybutyric acid.

The pharmaceutical compositions to be used for in vivo administrationmust be sterile. This is readily accomplished by, for example,filtration through sterile filtration membranes. Therapeutic antibodycompositions are generally placed into a container having a sterileaccess port, for example, an intravenous solution bag or vial having astopper pierceable by a hypodermic injection needle.

The pharmaceutical compositions described herein can be in unit dosageforms such as tablets, pills, capsules, powders, granules, solutions orsuspensions, or suppositories, for oral, parenteral or rectaladministration, or administration by inhalation or insufflation.For preparing solid compositions such as tablets, the principal activeingredient can be mixed with a pharmaceutical carrier, e.g.,conventional tableting ingredients such as corn starch, lactose,sucrose, sorbitol, talc, stearic acid, magnesium stearate, dicalciumphosphate or gums, and other pharmaceutical diluents, e.g., water, toform a solid preformulation composition containing a homogeneous mixtureof a compound of the present invention, or a non-toxic pharmaceuticallyacceptable salt thereof. When referring to these preformulationcompositions as homogeneous, it is meant that the active ingredient isdispersed evenly throughout the composition so that the composition maybe readily subdivided into equally effective unit dosage forms such astablets, pills and capsules. This solid preformulation composition isthen subdivided into unit dosage forms of the type described abovecontaining from 0.1 to about 500 mg of the active ingredient of thepresent invention. The tablets or pills of the novel composition can becoated or otherwise compounded to provide a dosage form affording theadvantage of prolonged action. For example, the tablet or pill cancomprise an inner dosage and an outer dosage component, the latter beingin the form of an envelope over the former. The two components can beseparated by an enteric layer that serves to resist disintegration inthe stomach and permits the inner component to pass intact into theduodenum or to be delayed in release. A variety of materials can be usedfor such enteric layers or coatings, such materials including a numberof polymeric acids and mixtures of polymeric acids with such materialsas shellac, cetyl alcohol and cellulose acetate. Suitable surface-activeagents include, in particular, non-ionic agents, such aspolyoxyethylenesorbitans (e.g., Tween™ 20, 40, 60, 80 or 85) and othersorbitans (e.g., Span™ 20, 40, 60, 80 or 85). Compositions with asurface-active agent will conveniently comprise between 0.05 and 5%surface-active agent, and can be between 0.1 and 2.5%. It will beappreciated that other ingredients may be added, for example mannitol orother pharmaceutically acceptable vehicles, if necessary.

Suitable emulsions may be prepared using commercially available fatemulsions, such as Intralipid™, Liposyn™, Infonutrol™, Lipofundin™ andLipiphysan™. The active ingredient may be either dissolved in apre-mixed emulsion composition or alternatively it may be dissolved inan oil (e.g., soybean oil, safflower oil, cottonseed oil, sesame oil,corn oil or almond oil) and an emulsion formed upon mixing with aphospholipid (e.g. egg phospholipids, soybean phospholipids or soybeanlecithin) and water. It will be appreciated that other ingredients maybe added, for example glycerol or glucose, to adjust the tonicity of theemulsion. Suitable emulsions will typically contain up to 20% oil, forexample, between 5 and 20%. The fat emulsion can comprise fat dropletsbetween 0.1 and 1.0 .im, particularly 0.1 and 0.5 .im, and have a pH inthe range of 5.5 to 8.0.

The emulsion compositions can be those prepared by mixing an antibodywith Intralipid™ or the components thereof (soybean oil, eggphospholipids, glycerol and water).

Pharmaceutical compositions for inhalation or insufflation includesolutions and suspensions in pharmaceutically acceptable, aqueous ororganic solvents, or mixtures thereof, and powders. The liquid or solidcompositions may contain suitable pharmaceutically acceptable excipientsas set out above. In some embodiments, the compositions are administeredby the oral or nasal respiratory route for local or systemic effect.

Compositions in preferably sterile pharmaceutically acceptable solventsmay be nebulized by use of gases. Nebulized solutions may be breatheddirectly from the nebulizing device or the nebulizing device may beattached to a face mask, tent or intermittent positive pressurebreathing machine. Solution, suspension or powder compositions may beadministered, preferably orally or nasally, from devices which deliverthe formulation in an appropriate manner.

Therapeutic Applications

The present disclosure provides methods of eliminating pathologic cellsexpressing Galectin-9, the method comprising administering to a subjecthaving pathologic cells expressing Galectin-9 an effective amount of apharmaceutical composition comprising an anti-Galectin-9 antibodydescribed herein. The present disclosure also provides methods ofinhibiting Galectin-9-mediated cell signaling in a subject, the methodcomprising administering to a subject in need thereof an effectiveamount of a pharmaceutical composition comprising an anti-Galectin-9antibody described herein.

To practice the methods disclosed herein, an effective amount of thepharmaceutical composition described herein can be administered to asubject (e.g., a human) in need of the treatment via a suitable route,systemically or locally. In some embodiments, the anti-Galectin-9antibodies are administered by intravenous administration, e.g., as abolus or by continuous infusion over a period of time, by intramuscular,intraperitoneal, intracerebrospinal, subcutaneous, intra-arterial,intra-articular, intrasynovial, intrathecal, intratumoral, oral,inhalation or topical routes. Commercially available nebulizers forliquid formulations, including jet nebulizers and ultrasonic nebulizersare useful for administration. Liquid formulations can be directlynebulized and lyophilized powder can be nebulized after reconstitution.Alternatively, the antibodies as described herein can be aerosolizedusing a fluorocarbon formulation and a metered dose inhaler, or inhaledas a lyophilized and milled powder.

The subject to be treated by the methods described herein can be amammal, more preferably a human. Mammals include, but are not limitedto, farm animals, sport animals, pets, primates, horses, dogs, cats,mice and rats. A human subject who needs the treatment may be a humanpatient having, at risk for, or suspected of having a targetdisease/disorder, such as a solid tumor, hematological malignancy,autoimmune disease (such as an allergic disorder), microbial disease,and fibrotic condition.

Examples of solid tumor cancers include pancreatic duct adenocarcinoma(PDA), colorectal cancer (CRC), melanoma, cholangiocarcinoma, breastcancer, lung cancer (for example, non-small cell lung cancer, NSCLC, andsmall cell lung cancer, SCLC), upper and lower gastrointestinalmalignancies (including, but not limited to, esophageal, gastric, andhepatobiliary cancer), squamous cell head and neck cancer, genitourinarycancers, ovarian cancer, and sarcomas. Hematological malignanciesinclude acute lymphoblastic leukemia, chronic lymphocytic leukemia,lymphomas, multiple myeloma, acute myelogenous leukemia, chronicmyelogenous leukemia, myelodysplastic syndromes and themyeloproliferative neoplasms, such as essential thrombocythemia,polycythemia vera and myelofibrosis. A subject having a solid tumor or ahematological malignancy can be identified by routine medicalexamination, e.g., laboratory tests, organ functional tests, CT scans,or ultrasounds. In some embodiments, the subject to be treated by themethod described herein may be a human cancer patient who has undergoneor is subjecting to an anti-cancer therapy, for example, chemotherapy,radiotherapy, immunotherapy, or surgery.

Examples of autoimmune diseases include rheumatoid conditions, metabolicand endocrine conditions, as well as respiratory and allergicconditions. A subject having an autoimmune disease can be identified byroutine medical examination, e.g., with laboratory tests, such asantinuclear antibodies, anti-mitochondrial autoantibodies,anti-neutrophil cytoplasmic antibody, anti-phospholipid antibodies,anti-citrullinated peptide (anti-CCP), anti-rheumatoid factor,immunoglobulin A, C-reactive protein test, complement test, erythrocytesedimentation rate (ESR) test, blood clotting profie, and proteinelectrophoresis/immunofixation electrophoresis, among others. In someembodiments, the subject to be treated by the method described hereinmay be a human subject with an autoimmune disease who has undergone oris subjecting to an autoimmune disease treatment, for example,immunosuppressive mediation, hormone replacement therapy, bloodtransfusions, anti-inflammatory medication, and/or pain medication.

Microbial diseases can be caused by a variety of pathogens, includingbacteria, fungi, protozoa and viruses. Exemplary infectious bacteriainclude Streptococcus pyogenes, Streptococcus pneumoniae, Neisseriagonorrheae, Neisseria meningitidis, Corynebacterium diphtherias,Clostridium botulinum, Clostridium perfringens, Clostridium tetani,Hemophilus influenzae, Klebsiella pneumoniae, Klebsiella ozaenas,Klebsiella rhinoscleromotis, Staphylococcus aureus, Vibrio colerae,Escherichia coli, Pseudomonas aeruginosa, Campylobacter (Vibrio) fetus,Aeromonas hydrophila, Bacillus aureus, Edwardsiella tarda, Yersiniaenterocolitica, Yersinia pestis, Yersinia pseudotuberculosis, Shigelladysenteriae, Shigella flexneri, Shigella sonnei, Salmonella typhimurium,Treponema pallidum, Treponema pertenue, Treponema carateneum, Borreliavincentii, Borrelia burgdorferi, Leptospira icterohemorrhagiae,Mycobacterium tuberculosis, Pneumocystis carinii, Francisellatularensis, Brucella abortus, Brucella suis, Brucella melitensis,Mycoplasma spp., Rickettsia prowazeki, Rickettsia tsutsugumushi, andChlamydia spp. Examples of pathologic fungi include Coccidioidesimmitis, Aspergillusfumigatus, Candida albicans, Blastomycesdermatitidis, Cryptococcus neoformans, and Histoplasma capsulatum.Pathologic protozoa include Entomoeba histolytica, Toxoplasma gondii,Trichomonas tenas, Trichomonas hominis, Trichomonas vaginalis,Tryoanosoma gambiense, Trypanosoma rhodesiense, Trypanosoma cruzi,Leishmania donovani, Leishmania tropica, Leishmania braziliensis,Pneumocystis pneumonia, Plasmodium vivax, Plasmodium falciparum, andPlasmodium malaria. Examples of helminiths include Enterobiusvermicularis, Trichuris trichiura, Ascaris lumbricoides, Trichinellaspiralis, Strongyloides stercoralis, Schistosoma japonicum, Schistosomamansoni, Schistosoma haematobium, and hookworms. Viral infectiousdiseases include those caused by Adenovirus, Lassa fever virus(Arenavirus), Astrovirus, Hantavirus, Rift Valley Fever virus(Phlebovirus), Calicivirus, Ebola virus, Marburg Virus, Japaneseencephalitis virus, Dengue virus, Yellow fever virus, Hepatitis C virus,Hepatitis G virus, Hepatitis B virus, Hepatitis D virus, Herpes simplexvirus 1, Herpes simplex virus 2, Cytomegalovirus, Epstein Barr virus,Varicella Zoster Virus, Human Herpesvirus 7, Human Herpesvirus 8,Influenza virus, Parainfluenza virus, Rubella virus, Mumps virus,Morbillivirus, Measles virus, Respiratory Syncytial virus,Papillomaviruses, JC virus (Polyomavirus), BK virus (Polyomavirus),Parvovirus, Coxsackie virus (A and B), Hepatitis A virus, Polioviruses,Rhinoviruses, Reovirus, Rabies Virus (Lyssavirus), HumanImmunodeficiency virus 1 and 2, and Human T-cell Leukemia virus. Asubject having a microbial disease can be identified by routine medicalexamination, e.g., laboratory tests. For example, microscopy (e.g.,Gram-positive and/or Gram-negative staining), sample culturing,biochemical tests (e.g., tests for metabolic and/or enzymatic products,such as fermentation products, acids, alcohol, or gases), and moleculardiagnostics (e.g., PCR) may be used. In some embodiments, the subject tobe treated by the method described herein may be a human infectiousdisease patient who has undergone or is subjecting to an antimicrobialtherapy, for example, immunotherapy.

Examples of fibrotic conditions include pulmonary fibrosis (e.g., cysticfibrosis, idiopathic pulmonary fibrosis), cirrhosis, biliary atresia,atrial fibrosis, endomyocardial fibrosis, glial scar, arthrofibrosis,Crohn's disease, Dupuytren's contracture, keloid, mediastinal fibrosis,myelofibrosis, nephrogenic systemic fibrosis, progressive massivefibrosis, retroperitoneal fibrosis, and scleroderma/systemic sclerosis.A subject having a fibrotic condition can be identified by routinemedical examination, e.g., laboratory tests, CT scans, X-rays,echocardiograms, or ultrasounds. In some embodiments, the subject to betreated by the method described herein may be a human fibrotic patientwho has undergone or is subjecting to an anti-fibrotic therapy, forexample medication, physical therapy, oxygen therapy, or surgery.

A subject suspected of having any of such target disease/disorder mightshow one or more symptoms of the disease/disorder. A subject at risk forthe disease/disorder can be a subject having one or more of the riskfactors for that disease/disorder.

As used herein, “an effective amount” refers to the amount of eachactive agent required to confer therapeutic effect on the subject,either alone or in combination with one or more other active agents. Insome embodiments, the therapeutic effect is reduced Galectin-9 activityand/or amount/expression, reduced Dectin-1 signaling, reduced TIM-3signaling, reduced CD206 signaling, or increased anti-tumor immuneresponses in the tumor microenvironment. Non-limiting examples ofincreased anti-tumor responses include increased activation levels ofeffector T cells, or switching of the TAMS from the M2 to the M1phenotype, and increased ADCC responses. Determination of whether anamount of the antibody achieved the therapeutic effect would be evidentto one of skill in the art. Effective amounts vary, as recognized bythose skilled in the art, depending on the particular condition beingtreated, the severity of the condition, the individual patientparameters including age, physical condition, size, gender and weight,the duration of the treatment, the nature of concurrent therapy (ifany), the specific route of administration and like factors within theknowledge and expertise of the health practitioner. These factors arewell known to those of ordinary skill in the art and can be addressedwith no more than routine experimentation. It is generally preferredthat a maximum dose of the individual components or combinations thereofbe used, that is, the highest safe dose according to sound medicaljudgment.

Empirical considerations, such as the half-life, generally willcontribute to the determination of the dosage. For example, antibodiesthat are compatible with the human immune system, such as humanizedantibodies or fully human antibodies, may be used to prolong half-lifeof the antibody and to prevent the antibody being attacked by the host'simmune system. Frequency of administration may be determined andadjusted over the course of therapy, and is generally, but notnecessarily, based on treatment and/or suppression and/or ameliorationand/or delay of a target disease/disorder. Alternatively, sustainedcontinuous release formulations of an antibody may be appropriate.Various formulations and devices for achieving sustained release areknown in the art.

In one example, dosages for an antibody as described herein may bedetermined empirically in individuals who have been given one or moreadministration(s) of the antibody. Individuals are given incrementaldosages of the antagonist. To assess efficacy of the antagonist, anindicator of the disease/disorder can be followed.

Generally, for administration of any of the antibodies described herein,such as those described in Table 1 or Table 2 herein, such as forexample, antibody 9.2-17 and antibody 9.1-8mut1, an initial candidatedosage can be about 2 mg/kg. For the purpose of the present disclosure,a typical daily dosage might range from about any of 0.1 μg/kg to 3μg/kg to 30 μg/kg to 300 μg/kg to 3 mg/kg, to 30 mg/kg to 100 mg/kg ormore, depending on the factors mentioned above. For repeatedadministrations over several days or longer, depending on the condition,the treatment is sustained until a desired suppression of symptomsoccurs or until sufficient therapeutic levels are achieved to alleviatea target disease or disorder, or a symptom thereof. An exemplary dosingregimen comprises administering an initial dose of about 2 mg/kg,followed by a weekly maintenance dose of about 1 mg/kg of the antibody,or followed by a maintenance dose of about 1 mg/kg every other week.However, other dosage regimens may be useful, depending on the patternof pharmacokinetic decay that the practitioner wishes to achieve. Forexample, dosing from one-four times a week is contemplated. In someembodiments, dosing ranging from about 3 μg/mg to about 2 mg/kg (such asabout 3 μg/mg, about 10 μg/mg, about 30 μg/mg, about 100 μg/mg, about300 μg/mg, about 1 mg/kg, and about 2 mg/kg) may be used. In someembodiments, dosing frequency is once every week, every 2 weeks, every 4weeks, every 5 weeks, every 6 weeks, every 7 weeks, every 8 weeks, every9 weeks, or every 10 weeks; or once every month, every 2 months, orevery 3 months, or longer. The progress of this therapy is easilymonitored by conventional techniques and assays. The dosing regimen(including the antibody used) can vary over time.

In some embodiments, for an adult patient of normal weight, dosesranging from about 0.3 to 5.00 mg/kg may be administered. In someexamples, the dosage of the anti-Galectin-9 antibody described hereincan be 10 mg/kg. The particular dosage regimen, i.e., dose, timing andrepetition, will depend on the particular individual and thatindividual's medical history, as well as the properties of theindividual agents (such as the half-life of the agent, and otherconsiderations well known in the art).

For the purpose of the present disclosure, the appropriate dosage of anantibody as described herein will depend on the specific antibody,antibodies, and/or non-antibody peptide (or compositions thereof)employed, the type and severity of the disease/disorder, whether theantibody is administered for preventive or therapeutic purposes,previous therapy, the patient's clinical history and response to theantagonist, and the discretion of the attending physician. Typically,the clinician will administer an antibody, until a dosage is reachedthat achieves the desired result. In some embodiments, the desiredresult is an increase in anti-tumor immune response in the tumormicroenvironment. Methods of determining whether a dosage resulted inthe desired result would be evident to one of skill in the art.Administration of one or more antibodies can be continuous orintermittent, depending, for example, upon the recipient's physiologicalcondition, whether the purpose of the administration is therapeutic orprophylactic, and other factors known to skilled practitioners. Theadministration of an antibody may be essentially continuous over apreselected period of time or may be in a series of spaced dose, e.g.,either before, during, or after developing a target disease or disorder.

As used herein, the term “treating” refers to the application oradministration of a composition including one or more active agents to asubject, who has a target disease or disorder, a symptom of thedisease/disorder, or a predisposition toward the disease/disorder, withthe purpose to cure, heal, alleviate, relieve, alter, remedy,ameliorate, improve, or affect the disorder, the symptom of the disease,or the predisposition toward the disease or disorder.

Alleviating a target disease/disorder includes delaying the developmentor progression of the disease, or reducing disease severity orprolonging survival. Alleviating the disease or prolonging survival doesnot necessarily require curative results. As used therein, “delaying”the development of a target disease or disorder means to defer, hinder,slow, retard, stabilize, and/or postpone progression of the disease.This delay can be of varying lengths of time, depending on the historyof the disease and/or individuals being treated. A method that “delays”or alleviates the development of a disease, or delays the onset of thedisease, is a method that reduces probability of developing one or moresymptoms of the disease in a given time frame and/or reduces extent ofthe symptoms in a given time frame, when compared to not using themethod. Such comparisons are typically based on clinical studies, usinga number of subjects sufficient to give a statistically significantresult.

“Development” or “progression” of a disease means initial manifestationsand/or ensuing progression of the disease. Development of the diseasecan be detectable and assessed using standard clinical techniques aswell known in the art. However, development also refers to progressionthat may be undetectable. For purpose of this disclosure, development orprogression refers to the biological course of the symptoms.“Development” includes occurrence, recurrence, and onset. As used herein“onset” or “occurrence” of a target disease or disorder includes initialonset and/or recurrence.

In some embodiments, the antibodies described herein, e.g., in Table 1and/or Table 2, including, but not limited to, 9.2-17 and 9.1-8mut13,are administered to a subject in need of the treatment at an amountsufficient to inhibit the activity of Galectin-9 (and/or Dectin-1 orTIM-3 or CD206) in immune suppressive immune cells in a tumor by atleast 20% (e.g., 30%, 40%, 50%, 60%, 70%, 80%, 90% or greater) in vivo.In other embodiments, the antibodies described herein, e.g., in Table 1and/or Table 2, including, but not limited to, 9.2-17 and 9.1-8mut13,are administered in an amount effective in reducing the activity levelof Galectin-9 (and/or Dectin-1 or TIM-3 or CD206) in immune suppressiveimmune cells in a tumor by at least 20% (e.g., 30%, 40%, 50%, 60%, 70%,80%, 90% or greater) (as compared to levels prior to treatment or in acontrol subject). In some embodiments, the antibodies described herein,e.g., in Table 1 and/or Table 2, including, but not limited to, 9.2-17and 9.1-8mut13, are administered to a subject in need of the treatmentat an amount sufficient to promote M1-like programming in TAMs by atleast 20% (e.g., 30%, 40%, 50%, 60%, 70%, 80%, 90% or greater) in vivo(as compared to levels prior to treatment or in a control subject).

In some embodiments, the antibodies described herein, e.g., in Table 1and/or Table 2, including, but not limited to, 9.2-17 and 9.1-8mut13,are administered to a subject in need thereof at an amount sufficient topromote ADCC in target cells in a tumor. In some embodiments, theantibodies described herein, e.g., in Table 1 and/or Table 2, including,but not limited to, 9.2-17 and 9.1-8mut13, are administered to a subjectin need of the treatment at an amount sufficient to promote ADCC intarget cells in a tumor by at least 20% (e.g., 30%, 40%, 50%, 60%, 70%,80%, 90% or greater) in vivo (as compared to levels prior to treatmentor in a control subject).

In some embodiments, the antibodies described herein, e.g., in Table 1and/or Table 2, including, but not limited to, 9.2-17 and 9.1-8mut13,are administered to a subject in need of the treatment at an amountsufficient to promote CDC in target cells in a tumor. In someembodiments, the antibodies described herein, e.g., in Table 1 and/orTable 2, including, but not limited to, 9.2-17 and 9.1-8mut13, areadministered to a subject in need of the treatment at an amountsufficient to promote CDC in target cells in a tumor by at least 20%(e.g., 30%, 40%, 50%, 60%, 70%, 80%, 90% or greater) in vivo (ascompared to levels prior to treatment or in a control subject).

In some embodiments, the antibodies described herein, e.g., in Table 1and/or Table 2, including, but not limited to, 9.2-17 and 9.1-8mut13,are administered to a subject in need of the treatment at an amountsufficient to promote ADCP in target cells in a tumor. In someembodiments, the antibodies described herein, e.g., in Table 1 and/orTable 2, including, but not limited to, 9.2-17 and 9.1-8mut13, areadministered to a subject in need of the treatment at an amountsufficient to promote ADCP in target cells in a tumor by at least 20%(e.g., 30%, 40%, 50%, 60%, 70%, 80%, 90% or greater) in vivo (ascompared to levels prior to treatment or in a control subject).

In some embodiments, the antibodies described herein, e.g., in Table 1and/or Table 2, including, but not limited to, 9.2-17 and 9.1-8mut13,are administered to a subject in need of the treatment at an amountsufficient to promote T cell activation in a tumor. In some embodiments,the antibodies described herein, e.g., in Table 1 and/or Table 2,including, but not limited to, 9.2-17 and 9.1-8mut13, are administeredto a subject in need of the treatment at an amount sufficient to promoteT cell activation in a tumor by at least 20% (e.g., 30%, 40%, 50%, 60%,70%, 80%, 90% or greater) in vivo (as compared to levels prior totreatment or in a control subject).

In some embodiments, the antibodies described herein, e.g., in Table 1and/or Table 2, including, but not limited to, 9.2-17 and 9.1-8mut13,are administered to a subject in need of the treatment at an amountsufficient to promote CD4+ T cell activation in a tumor. In someembodiments, the antibodies described herein, e.g., in Table 1 and/orTable 2, including, but not limited to, 9.2-17 and 9.1-8mut13, areadministered to a subject in need of the treatment at an amountsufficient to promote CD4+ T cell activation in a tumor by at least 20%(e.g., 30%, 40%, 50%, 60%, 70%, 80%, 90% or greater) in vivo (ascompared to levels prior to treatment or in a control subject).

In some embodiments, the antibodies described herein, e.g., in Table 1and/or Table 2, including, but not limited to, 9.2-17 and 9.1-8mut13,are administered to a subject in need of the treatment at an amountsufficient to induce CD44 expression in CD4+ cells in a tumor. In someembodiments, the antibodies described herein, e.g., in Table 1 and/orTable 2, including, but not limited to, 9.2-17 and 9.1-8mut13, areadministered to a subject in need of the treatment at an amountsufficient to induce CD44 expression in CD4+ cells in a tumor by atleast 20% (e.g., 30%, 40%, 50%, 60%, 70%, 80%, 90% or greater) in vivo(as compared to levels prior to treatment or in a control subject).

In some embodiments, the antibodies described herein, e.g., in Table 1and/or Table 2, including, but not limited to, 9.2-17 and 9.1-8mut13,are administered to a subject in need of the treatment at an amountsufficient to induce TNFalpha expression in CD4+ cells in a tumor. Insome embodiments, the antibodies described herein, e.g., in Table 1and/or Table 2, including, but not limited to, 9.2-17 and 9.1-8mut13,are administered to a subject in need of the treatment at an amountsufficient to induce TNFalpha expression in CD4+ cells in a tumor by atleast 20% (e.g., 30%, 40%, 50%, 60%, 70%, 80%, 90% or greater) in vivo(as compared to levels prior to treatment or in a control subject).

In some embodiments, the antibodies described herein, e.g., in Table 1and/or Table 2, including, but not limited to, 9.2-17 and 9.1-8mut13,are administered to a subject in need of the treatment at an amountsufficient to promote CD8+ T cell activation in a tumor. In someembodiments, the antibodies described herein, e.g., in Table 1 and/orTable 2, including, but not limited to, 9.2-17 and 9.1-8mut13, areadministered to a subject in need of the treatment at an amountsufficient to promote CD8+ T cell activation in a tumor by at least 20%(e.g., 30%, 40%, 50%, 60%, 70%, 80%, 90% or greater) in vivo (ascompared to levels prior to treatment or in a control subject).

In some embodiments, the antibodies described herein, e.g., in Table 1and/or Table 2, including, but not limited to, 9.2-17 and 9.1-8mut13,are administered to a subject in need of the treatment at an amountsufficient to induce CD44 expression in CD8+ cells in a tumor. In someembodiments, the antibodies described herein, e.g., in Table 1 and/orTable 2, including, but not limited to, 9.2-17 and 9.1-8mut13, areadministered to a subject in need of the treatment at an amountsufficient to induce CD44 expression in CD8+ cells in a tumor by atleast 20% (e.g., 30%, 40%, 50%, 60%, 70%, 80%, 90% or greater) in vivo(as compared to levels prior to treatment or in a control subject).

In some embodiments, the antibodies described herein, e.g., in Table 1and/or Table 2, including, but not limited to, 9.2-17 and 9.1-8mut13,are administered to a subject in need of the treatment at an amountsufficient to induce TNFalpha expression in CD8+ cells in a tumor. Insome embodiments, the antibodies described herein, e.g., in Table 1and/or Table 2, including, but not limited to, 9.2-17 and 9.1-8mut13,are administered to a subject in need of the treatment at an amountsufficient to induce TNFalpha expression in CD8+ cells in a tumor by atleast 20% (e.g., 30%, 40%, 50%, 60%, 70%, 80%, 90% or greater) in vivo(as compared to levels prior to treatment or in a control subject).

In any of these embodiments, the antibodies described herein, e.g., inTable 1 and/or Table 2, including, but not limited to, 9.2-17 and9.1-8mut13, are administered to a subject having cancer, wherein thecancer is selected from pancreatic cancer, e.g., pancreatic ductaladenocarcinoma, cholangiocarcinoma, hepatocellular carcinoma, colorectalcancer, melanoma, renal cell carcinoma, and acute myeloid leukemia.

An analysis was conducted by the inventors using TCGA (The Cancer GenomeAtlas) RNA Seq data from 29 different types of solid tumors utilizingapproximately 40,000 individual samples, to analyze the correlationbetween T cell infiltration in a particular tumor type and levels ofPD1, PD-L1, IFNgamma and TNF alpha expression. Tumor types were rankedby median expression for a given gene. Gene expression for 4 TCRcomponents (CD3d, CD3e, CD3g, CD3z) and the T cell specific effectorprotein kinase ZAP70 were used to establish the relative level of T cellassociation with a given tumor type. TBX21, a Th1 cell-specifictranscription factor that controls the expression of the hallmark Th1cytokines interferon-gamma (IFNg) and tumor necrosis factor (TNF) wasalso evaluated.

According to the analysis, T cell association levels (surrogate for Tcell infiltration) with individual tumor types is generally proportionalto the ranking of IFNg expression with the exception of pancreaticcancer (PDA). In PDA the level of IFNg transcription is significantlysuppressed suggesting over that observed in other solid tumors implyingthat the immunosuppressive environment of PDA is particularly robust. Inthis dataset, the expression of TNF does not generally correlate withthe degree of T cell infiltration. Of note, under most conditions TNF isproduced by activated macrophages with less contribution by Th1 T cells,NK cells, neutrophils, mast cells, and eosinophils.

Without wishing to be bound by theory, increasing levels of IFNgamma maybe particularly useful to combat the immunosuppressive environment andre-activate myeloid and lymphoid response particularly in PDA.Accordingly, in one embodiment, methods of increasing levels of IFNgammain a cancer are provided herein, wherein the method comprisesadministering an anti-Galectin-9 antibody, e.g., as described herein inTable 1 and/or Table 2, including, but not limited to, antibody 9.1-8m13and/or antibody 9.2-17, and wherein the levels of IFNgamma in the cancerare low prior prior the administration, e.g., as assessed relative tolevels of expression of T cell markers. In some embodiments the canceris PDA.

Accordingly, in some embodiments, methods of increasing levels ofIFNgamma in a cancer are provided herein, wherein the method comprisesadministering an anti-Galectin-9 antibody, e.g., as described herein inTable 1 and/or Table 2, including, but not limited to, antibody 9.1-8m13and/or antibody 9.2-17, and wherein the cancer is PDA. In someembodiments, methods are provided herein, wherein the antibodiesdescribed herein, e.g., in Table 1 and/or Table 2, including, but notlimited to, 9.2-17 and 9.1-8mut13, are administered to a subject in needof the treatment at an amount sufficient to induce IFNgamma expressionin effector T cells in a tumor. In some embodiments, the antibodiesdescribed herein, e.g., in Table 1 and/or Table 2, including, but notlimited to, 9.2-17 and 9.1-8mut13, are administered to a subject in needof the treatment at an amount sufficient to induce IFNgamma expressionin effector T cells in a tumor. In In some embodiments, the antibodiesdescribed herein, e.g., in Table 1 and/or Table 2, including, but notlimited to, 9.2-17 and 9.1-8mut13, are administered to a subject in needof the treatment at an amount sufficient to induce IFNgamma expressionin CD4+ cells in a tumor by at least 20% (e.g., 30%, 40%, 50%, 60%, 70%,80%, 90% or greater) in vivo (as compared to levels prior to treatmentor in a control subject).

In some embodiments, methods are provided herein, wherein the antibodiesdescribed herein, e.g., in Table 1 and/or Table 2, including, but notlimited to, 9.2-17 and 9.1-8mut13, are administered to a subject in needof the treatment at an amount sufficient to induce IFNgamma expressionin CD4+ cells in a tumor. In some embodiments, the antibodies describedherein, e.g., in Table 1 and/or Table 2, including, but not limited to,9.2-17 and 9.1-8mut13, are administered to a subject in need of thetreatment at an amount sufficient to induce IFNgamma expression in CD4+cells in a tumor. In some embodiments, the antibodies described herein,e.g., in Table 1 and/or Table 2, including, but not limited to, 9.2-17and 9.1-8mut13, are administered to a subject in need of the treatmentat an amount sufficient to induce IFNgamma expression in CD4+ cells in atumor by at least 20% (e.g., 30%, 40%, 50%, 60%, 70%, 80%, 90% orgreater) in vivo (as compared to levels prior to treatment or in acontrol subject).

In some embodiments, methods are provided herein, wherein the antibodiesdescribed herein, e.g., in Table 1 and/or Table 2, including, but notlimited to, 9.2-17 and 9.1-8mut13, are administered to a subject in needof the treatment at an amount sufficient to induce IFNgamma expressionin CD8+ cells in a tumor. In some embodiments, the antibodies describedherein, e.g., in Table 1 and/or Table 2, including, but not limited to,9.2-17 and 9.1-8mut13, are administered to a subject in need of thetreatment at an amount sufficient to induce IFNgamma expression in CD8+cells in a tumor. In some embodiments, the antibodies described herein,e.g., in Table 1 and/or Table 2, including, but not limited to, 9.2-17and 9.1-8mut13, are administered to a subject in need of the treatmentat an amount sufficient to induce IFNgamma expression in CD8+ cells in atumor by at least 20% (e.g., 30%, 40%, 50%, 60%, 70%, 80%, 90% orgreater) in vivo (as compared to levels prior to treatment or in acontrol subject).

In some embodiments, methods are provided herein, wherein immune cellpopulations in tumor samples are analyzed in vitro or ex vivo.Accordingly methods are provided herein, wherein the antibodiesdescribed herein, e.g., in Table 1 and/or Table 2, including, but notlimited to, 9.2-17 and 9.1-8mut13, are provided in vitro or ex vivo atan amount sufficient to induce IFNgamma expression in effector T cellsin a tumor. In some embodiments, the antibodies described herein, e.g.,in Table 1 and/or Table 2, including, but not limited to, 9.2-17 and9.1-8mut13, are provided in vitro or ex vivo at an amount sufficient toinduce IFNgamma expression in effector T cells in a tumor. In someembodiments, the antibodies described herein, e.g., in Table 1 and/orTable 2, including, but not limited to, 9.2-17 and 9.1-8mut13, induceIFNgamma expression in CD4+ cells in a tumor by at least 20% (e.g., 30%,40%, 50%, 60%, 70%, 80%, 90% or greater) in vitro or ex vivo. In someembodiments, administration of one or more of the antibodies describedherein results in a reduction in tumor size, reduction in tumor growth,elimination of the tumor, reduction in number of metastatic lesions overtime, complete response, partial response, or stable disease.Conventional methods, known to those of ordinary skill in the art ofmedicine, can be used to administer the pharmaceutical composition tothe subject, depending upon the type of disease to be treated or thesite of the disease. This composition can also be administered via otherconventional routes, e.g., administered orally, parenterally, byinhalation spray, topically, rectally, nasally, buccally, vaginally orvia an implanted reservoir. The term “parenteral” as used hereinincludes subcutaneous, intracutaneous, intravenous, intramuscular,intraarticular, intraarterial, intrasynovial, intrasternal, intrathecal,intralesional, intratumoral, and intracranial injection or infusiontechniques. In addition, it can be administered to the subject viainjectable depot routes of administration such as using 1-, 3-, or6-month depot injectable or biodegradable materials and methods. In someexamples, the pharmaceutical composition is administered intraocularlyor intravitreally.

Injectable compositions may contain various carriers such as vegetableoils, dimethylactamide, dimethyformamide, ethyl lactate, ethylcarbonate, isopropyl myristate, ethanol, and polyols (glycerol,propylene glycol, liquid polyethylene glycol, and the like). Forintravenous injection, water soluble antibodies can be administered bythe drip method, whereby a pharmaceutical formulation containing theantibody and a physiologically acceptable excipient is infused.Physiologically acceptable excipients may include, for example, 5%dextrose, 0.9% saline, Ringer's solution or other suitable excipients.Intramuscular preparations, e.g., a sterile formulation of a suitablesoluble salt form of the antibody, can be dissolved and administered ina pharmaceutical excipient such as Water-for-Injection, 0.9% saline, or5% glucose solution.

In one embodiment, an antibody is administered via site-specific ortargeted local delivery techniques. Examples of site-specific ortargeted local delivery techniques include various implantable depotsources of the antibody or local delivery catheters, such as infusioncatheters, an indwelling catheter, or a needle catheter, syntheticgrafts, adventitial wraps, shunts and stents or other implantabledevices, site specific carriers, direct injection, or directapplication. See, e.g., PCT Publication No. WO 00/53211 and U.S. Pat.No. 5,981,568.

Targeted delivery of therapeutic compositions containing an antisensepolynucleotide, expression vector, or subgenomic polynucleotides canalso be used. Receptor-mediated DNA delivery techniques are describedin, for example, Findeis et al., Trends Biotechnol. (1993) 11:202; Chiouet al., Gene Therapeutics: Methods And Applications Of Direct GeneTransfer (J. A. Wolff, ed.) (1994); Wu et al., J. Biol. Chem. (1988)263:621; Wu et al., J. Biol. Chem. (1994) 269:542; Zenke et al., Proc.Natl. Acad. Sci. USA (1990) 87:3655; Wu et al., J. Biol. Chem. (1991)266:338.

Therapeutic compositions containing a polynucleotide (e.g., thoseencoding the antibodies described herein) are administered in a range ofabout 100 ng to about 200 mg of DNA for local administration in a genetherapy protocol. In some embodiments, concentration ranges of about 500ng to about 50 mg, about 1 μg to about 2 mg, about 5 μg to about 500 μg,and about 20 μg to about 100 μg of DNA or more can also be used during agene therapy protocol.

The therapeutic polynucleotides and polypeptides described herein can bedelivered using gene delivery vehicles. The gene delivery vehicle can beof viral or non-viral origin (see generally, Jolly, Cancer Gene Therapy(1994) 1:51; Kimura, Human Gene Therapy (1994) 5:845; Connelly, HumanGene Therapy (1995) 1:185; and Kaplitt, Nature Genetics (1994) 6:148).Expression of such coding sequences can be induced using endogenousmammalian or heterologous promoters and/or enhancers. Expression of thecoding sequence can be either constitutive or regulated.

Viral-based vectors for delivery of a desired polynucleotide andexpression in a desired cell are well known in the art. Exemplaryviral-based vehicles include, but are not limited to, recombinantretroviruses (see, e.g., PCT Publication Nos. WO 90/07936; WO 94/03622;WO 93/25698; WO 93/25234; WO 93/11230; WO 93/10218; WO 91/02805; U.S.Pat. Nos. 5,219,740 and 4,777,127; GB Patent No. 2,200,651; and EPPatent No. 0 345 242), alphavirus-based vectors (e.g., Sindbis virusvectors, Semliki forest virus (ATCC VR-67; ATCC VR-1247), Ross Rivervirus (ATCC VR-373; ATCC VR-1246) and Venezuelan equine encephalitisvirus (ATCC VR-923; ATCC VR-1250; ATCC VR 1249; ATCC VR-532)), andadeno-associated virus (AAV) vectors (see, e.g., PCT Publication Nos. WO94/12649, WO 93/03769; WO 93/19191; WO 94/28938; WO 95/11984 and WO95/00655). Administration of DNA linked to killed adenovirus asdescribed in Curiel, Hum. Gene Ther. (1992) 3:147 can also be employed.

Non-viral delivery vehicles and methods can also be employed, including,but not limited to, polycationic condensed DNA linked or unlinked tokilled adenovirus alone (see, e.g., Curiel, Hum. Gene Ther. (1992)3:147); ligand-linked DNA (see, e.g., Wu, J. Biol. Chem. (1989)264:16985); eukaryotic cell delivery vehicles cells (see, e.g., U.S.Pat. No. 5,814,482; PCT Publication Nos. WO 95/07994; WO 96/17072; WO95/30763; and WO 97/42338) and nucleic charge neutralization or fusionwith cell membranes. Naked DNA can also be employed. Exemplary naked DNAintroduction methods are described in PCT Publication No. WO 90/11092and U.S. Pat. No. 5,580,859. Liposomes that can act as gene deliveryvehicles are described in U.S. Pat. No. 5,422,120; PCT Publication Nos.WO 95/13796; WO 94/23697; WO 91/14445; and EP Patent No. 0524968.Additional approaches are described in Philip, Mol. Cell. Biol. (1994)14:2411, and in Woffendin, Proc. Natl. Acad. Sci. (1994) 91:1581.

The particular dosage regimen, i.e., dose, timing and repetition, usedin the method described herein will depend on the particular subject andthat subject's medical history.

In some embodiments, more than one antibody, or a combination of anantibody and another suitable therapeutic agent, may be administered toa subject in need of the treatment. The antibody can also be used inconjunction with other agents that serve to enhance and/or complementthe effectiveness of the agents. Treatment efficacy for a targetdisease/disorder can be assessed by methods well-known in the art.

In some embodiments, the disclosure provides a method for suppressingDectin-1 signaling e.g., in immune suppressive immune cells, e.g., tumorinfiltrating immune cells, such as macrophages, the method comprisingproviding or administering an anti-Galectin-9 antibody described herein,e.g., in Table 1 and/or Table 2, or antigen binding fragment thereof toa subject. In some embodiments, the anti-Galectin-9 antibody is a9.1-8mut13 antibody and/or a 9.2-17 antibody. In some embodiments, themethod suppresses Dectin-1 signaling, e.g., in immune suppressive immunecells, by at least 30% (e.g., 31%, 35%, 40%, 50%, 60%, 70%, 80%, 90%,95% or greater, including any increment therein).

In some embodiments, the disclosure provides a method for suppressingTIM-3 signaling e.g., in tumor infiltrating immune cells, the methodcomprising providing or administering an anti-Galectin-9 antibodydescribed herein, e.g., in Table 1 and/or Table 2, or antigen bindingfragment thereof to a subject. In some embodiments, the anti-Galectin-9antibody is a 9.1-8mut13 antibody and/or a 9.2-17 antibody. In someembodiments, the method suppresses the TIM-3 signaling, e.g., in tumorinfiltrating immune cells by at least 30% (e.g., 31%, 35%, 40%, 50%,60%, 70%, 80%, 90%, 95% or greater, including any increment therein).

In some embodiments, the disclosure provides a method for suppressingCD206 signaling, e.g., in tumor infiltrating immune cells, e.g., inmacrophages, the method comprising providing or administering ananti-Galectin-9 antibody described herein, e.g., in Table 1 and/or Table2, or antigen binding fragment thereof to a subject. In someembodiments, the anti-Galectin-9 antibody is a 9.1-8mut13 antibodyand/or a 9.2-17 antibody. In some embodiments, the method suppresses theCD206 signaling, e.g., in tumor infiltrating immune cells, e.g.,macrophages, by at least 30% (e.g., 31%, 35%, 40%, 50%, 60%, 70%, 80%,90%, 95% or greater, including any increment therein).

In some embodiments, the disclosure provides a method for inducing cellcytotoxicity, such as ADCC, in target cells expressing Galectin-9, e.g.,wherein the target cells are cancer cells or immune suppressive immunecells, the method comprising providing or administering ananti-Galectin-9 antibody described herein, e.g., in Table 1 and/or Table2, or antigen binding fragment thereof to a subject. In someembodiments, the anti-Galectin-9 antibody is a 9.1-8mut13 antibodyand/or a 9.2-17 antibody. In some embodiments, the method inducesapoptosis in immune cells such as T cells by at least 30% (e.g., 31%,35%, 40%, 50%, 60%, 70%, 80%, 90%, 95% or greater, including anyincrement therein).

In some embodiments, the disclosure provides a method for inducing cellcytotoxicity such as complement-dependent cytotoxicity (CDC) againsttarget cells expressing Galectin-9 in a subject, the method comprisingproviding or administering an anti-Galectin-9 antibody described herein,e.g., in Table 1 and/or Table 2, or antigen binding fragment thereof toa subject. In some embodiments, the anti-Galectin-9 antibody is a9.1-8mut13 antibody and/or a 9.2-17 antibody. In some embodiments, themethod induces cell cytotoxicity such as complement-dependentcytotoxicity (CDC) against target cells expressing Galectin-9 at least30% (e.g., 31%, 35%, 40%, 50%, 60%, 70%, 80%, 90%, 95% or greater,including any increment therein).

In some embodiments, the disclosure provides a method for inducing cellcytotoxicity, such as ADCC, e.g., against target cells expressingGalectin-9 in a subject, the method comprising providing oradministering an anti-Galectin-9 antibody described herein, e.g., inTable 1 and/or Table 2, or antigen binding fragment thereof to asubject. In some embodiments, the anti-Galectin-9 antibody is a9.1-8mut13 antibody and/or a 9.2-17 antibody. In some embodiments, themethod induces cell cytotoxicity by at least 30% (e.g., 31%, 35%, 40%,50%, 60%, 70%, 80%, 90%, 95% or greater, including any incrementtherein).

In some embodiments, the disclosure provides a method for inducingphagocytosis of target cells expressing Galectin-9 (ADCP), the methodcomprising providing or administering an anti-Galectin-9 antibodydescribed herein, e.g., in Table 1 and/or Table 2, or antigen bindingfragment thereof to a subject. In some embodiments, the anti-Galectin-9antibody is a 9.1-8mut13 antibody and/or a 9.2-17 antibody. In someembodiments, the anti-Galectin-9 antibody increases phagocytosis oftarget cells by at least 30% (e.g., 31%, 35%, 40%, 50%, 60%, 70%, 80%,90%, 95% or greater, including any increment therein).

In some embodiments, the disclosure provides a method for inducing cellcytotoxicity such as complement-dependent cytotoxicity (CDC) againsttarget cells expressing Galectin-9, the method comprising providing oradministering an anti-Galectin-9 antibody described herein, e.g., inTable 1 and/or Table 2, or antigen binding fragment thereof to asubject. In some embodiments, the anti-Galectin-9 antibody is a9.1-8mut13 antibody and/or a 9.2-17 antibody. In some embodiments, themethod induces cell cytotoxicity against target cells by at least 30%(e.g., 31%, 35%, 40%, 50%, 60%, 70%, 80%, 90%, 95% or greater, includingany increment therein).

In some embodiments, the disclosure provides a method for inducing Tcell activation, e.g., in tumor infiltrating T cells, i.e., suppressGalectin-9 mediated inhibition of T cell activation, either directly orindirectly, the method comprising providing or administering ananti-Galectin-9 antibody described herein, e.g., in Table 1 and/or Table2, or antigen binding fragment thereof to a subject. In someembodiments, the anti-Galectin-9 antibody is a 9.1-8mut13 antibodyand/or a 9.2-17 antibody. In some embodiments, the method promotes Tcell activation by at least 30% (e.g., 31%, 35%, 40%, 50%, 60%, 70%,80%, 90%, 95% or greater, including any increment therein).

In some embodiments, the disclosure provides a method for promoting CD4+cell activation, the method comprising providing or administering ananti-Galectin-9 antibody described herein, e.g., in Table 1 and/or Table2, or antigen binding fragment thereof to a subject. In someembodiments, the anti-Galectin-9 antibody is a 9.1-8mut13 antibodyand/or a 9.2-17 antibody. In some embodiments, the method promotes CD4+cell activation by at least 30% (e.g., 31%, 35%, 40%, 50%, 60%, 70%,80%, 90%, 95% or greater, including any increment therein).

In some embodiments, the disclosure provides a method for inducing CD44expression in CD4+ cells, the method comprising providing oradministering an anti-Galectin-9 antibody described herein, e.g., inTable 1 and/or Table 2, or antigen binding fragment thereof to asubject. In some embodiments, the anti-Galectin-9 antibody is a9.1-8mut13 antibody and/or a 9.2-17 antibody. In some embodiments, themethod increases CD44 expression in CD4+ cells by at least 30% (e.g.,31%, 35%, 40%, 50%, 60%, 70%, 80%, 90%, 95% or greater, including anyincrement therein).

In some embodiments, the disclosure provides a method for inducingIFNgamma expression in CD4+ cells, the method comprising providing oradministering an anti-Galectin-9 antibody described herein, e.g., inTable 1 and/or Table 2, or antigen binding fragment thereof to asubject. In some embodiments, the anti-Galectin-9 antibody is a9.1-8mut13 antibody and/or a 9.2-17 antibody. In some embodiments, themethod increases IFNgamma expression in CD4+ cells by at least 30%(e.g., 31%, 35%, 40%, 50%, 60%, 70%, 80%, 90%, 95% or greater, includingany increment therein).

In some embodiments, the disclosure provides a method for inducingTNFalpha expression in CD4+ cells, the method comprising providing oradministering an anti-Galectin-9 antibody described herein, e.g., inTable 1 and/or Table 2, or antigen binding fragment thereof to asubject. In some embodiments, the anti-Galectin-9 antibody is a9.1-8mut13 antibody and/or a 9.2-17 antibody. In some embodiments, themethod increases TNFalpha expression in CD4+ cells by at least 30%(e.g., 31%, 35%, 40%, 50%, 60%, 70%, 80%, 90%, 95% or greater, includingany increment therein).

In some embodiments, the disclosure provides a method for inducing CD44expression in CD8+ cells, the method comprising providing oradministering an anti-Galectin-9 antibody described herein, e.g., inTable 1 and/or Table 2, or antigen binding fragment thereof to asubject. In some embodiments, the anti-Galectin-9 antibody is a9.1-8mut13 antibody and/or a 9.2-17 antibody. In some embodiments, themethod increases CD44 expression in CD8+ cells by at least 30% (e.g.,31%, 35%, 40%, 50%, 60%, 70%, 80%, 90%, 95% or greater, including anyincrement therein).

In some embodiments, the disclosure provides a method for inducingIFNgamma expression in CD8+ cells, the method comprising providing oradministering an effective amount of an anti-Galectin-9 antibodydescribed herein, e.g., in Table 1 and/or Table 2, or antigen bindingfragment thereof to a subject. In some embodiments, the anti-Galectin-9antibody is a 9.1-8mut13 antibody and/or a 9.2-17 antibody. In someembodiments, the method increases IFNgamma expression in CD8+ cells byat least 30% (e.g., 31%, 35%, 40%, 50%, 60%, 70%, 80%, 90%, 95% orgreater, including any increment therein).

In some embodiments, the disclosure provides a method for inducingTNFalpha expression in CD8+ cells, the method comprising providing oradministering an effective amount of an anti-Galectin-9 antibodydescribed herein, e.g., in Table 1 and/or Table 2, or antigen bindingfragment thereof to a subject. In some embodiments, the anti-Galectin-9antibody is a 9.1-8mut13 antibody and/or a 9.2-17 antibody. In someembodiments, the method increases TNFalpha expression in CD8+ cells byat least 30% (e.g., 31%, 35%, 40%, 50%, 60%, 70%, 80%, 90%, 95% orgreater, including any increment therein).

In some of these embodiments, the methods comprising providing oradministering an effective amount of an anti-Galectin-9 antibodydescribed herein, induce CD44, IFNgamma, and/or TNFalpha in CD4+ andCD8+ cells. The method embodiments described supra, for suppressingDectin-1 signaling, for suppressing TIM-3 signaling, for suppressingCD206 signaling, for inducing ADCC against target cells, for inducingCDC against target cell, for inducing ADCP against target cells, forinducing T cell activation, for promoting CD4+ cell activation, forinducing CD44 expression in CD4+ cells, for inducing IFNgamma expressionin CD4+ cells, for inducing TNFalpha expression in CD4+ cells, forinducing CD44 expression in CD8+ cells, for inducing IFNgamma expressionin CD8+ cells, method for inducing TNFalpha expression in CD8+ cells,wherein the method includes administering to a subject in need thereofan effective amount of an anti-Galectin-9 antibody described herein,e.g., in Table 1 and/or Table 2, or antigen binding fragment thereof.

The method embodiments described supra (for suppressing Dectin-1signaling, for suppressing TIM-3 signaling, for suppressing CD206signaling, for inducing ADCC against target cells, for inducing CDCagainst target cell, for inducing ADCP against target cells, forinducing T cell activation, for promoting CD4+ cell activation, forinducing CD44 expression in CD4+ cells, for inducing IFNgamma expressionin CD4+ cells, for inducing TNFalpha expression in CD4+ cells, forinducing CD44 expression in CD8+ cells, for inducing IFNgamma expressionin CD8+ cells, for inducing TNFalpha expression in CD8+ cells), whereinthe method includes providing an effective amount of an anti-Galectin-9antibody described herein, e.g., in Table 1 and/or Table 2, or antigenbinding fragment thereof, to a sample isolated from a tumor, andmeasuring in vitro or ex vivo one or more parameters selected fromDectin-1 suppression, for TIM-3 suppression, for CD206 suppression, ADCCinduction, CDC induction, ADCP induction, induction of T cellactivation, promotion of CD4+ cell activation, induction of CD44expression in CD4+ cells, induction of IFNgamma expression in CD4+cells, induction of TNFalpha expression in CD4+ cells, induction of CD44expression in CD8+ cells, induction of IFNgamma expression in CD8+cells, induction of TNFalpha expression in CD8+ cells.

The in vivo methods embodiments described supra, wherein the subject inneed of administration has cancer, and wherein the cancer is selectedfrom pancreatic cancer, e.g., pancreatic ductal adenocarcinoma,cholangiocarcinoma, hepatocellular carcinoma, colorectal cancer,melanoma, renal cell carcinoma, and acute myeloid leukemia. In someembodiments, cancer low levels of IFNgamma expression, relative toexpression of T cell markers. In some embodiments, the cancer is PDA.

The in vitro or ex vivo method embodiments described supra, wherein thesample isolated from a tumor is from a cancer selected from pancreaticcancer, e.g., pancreatic ductal adenocarcinoma, cholangiocarcinoma,hepatocellular carcinoma, colorectal cancer, melanoma, renal cellcarcinoma, and acute myeloid leukemia.

Combination Therapy

Any of the anti-Galectin-9 antibodies described herein may be utilizedin conjunction with other types of therapy for cancer or autoimmunediseases, such as chemotherapy, surgery, radiation, gene therapy, or inconjunction with other types of therapy for autoimmune diseases, such asimmunosuppressive mediation, hormone replacement therapy, bloodtransfusions, anti-inflammatory medication, and/or pain medication andso forth. Such therapies can be administered simultaneously orsequentially (in any order) with the immunotherapy according to thepresent disclosure.

In some embodiments, methods are provided herein, wherein theanti-Galectin-9 antibodies described herein are utilized in conjunctionwith other types of therapy for cancer or autoimmune diseases, such aschemotherapy, surgery, radiation, gene therapy, or in conjunction withother types of therapy for autoimmune diseases, such asimmunosuppressive mediation, hormone replacement therapy, bloodtransfusions, anti-inflammatory medication, and/or pain medication andso forth. In some embodiments, the methods include the steps ofadministering the anti-Galectin-9 antibodies, such as any of theanti-Galectin-9 antibodies described herein, e.g., in Table 1 and/orTable 2, simultaneously or sequentially (in any order) with theimmunotherapy according to the present disclosure. When co-administeredwith an additional therapeutic agent, suitable therapeutically effectivedosages for each agent may be lowered due to the additive action orsynergy.

In some embodiments, the methods are provided herein, wherein theanti-Galectin-9 antibody, for example antibody 9.2-17 or 9.1-8mut13, iscombined with other immunomodulatory treatments such as, e.g.,inhibitors of a checkpoint molecule (e.g., PD-1, PD-L1, PD-L2, CTLA-4,LAG3, TIM3, or A2aR), activators of a co-stimulatory receptor (e.g.,DX40, GITR, CD137, CD40, CD27, and ICOS), and/or inhibitors of an innateimmune cell target (e.g., KIR, NKG2A, CD96, TLR, and IDO). Without beingbound by theory, it is thought that anti-Galectin-9 antibodies, throughtheir inhibition of Dectin-1, can reprogram immune responses againsttumor cells via, e.g., inhibiting the activity of γδ T cells infiltratedinto tumor microenvironment, and/or enhancing immune surveillanceagainst tumor cells by, e.g., activating CD4+ and/or CD8+ T cells. Thus,combined use of an anti-Galectin-9 antibody and an immunomodulatoryagent such as those described herein would be expected to significantlyenhance anti-tumor efficacy.

In some embodiments, the methods are provided, wherein theanti-Galectin-9 antibody is administered concurrently with a checkpointinhibitor. In some embodiments, wherein the anti-Galectin-9 antibody isadministered before or after a checkpoint inhibitor. In someembodiments, the checkpoint inhibitor is administered systemically. Insome embodiments, the checkpoint inhibitor is administered locally.

In some embodiments, the methods are provided, wherein theanti-Galectin-9 antibody, such as any of the Galectin-9 antibodiesdescribed herein in Table 1 and/or Table 2, such as 9.2-17 or9.1-8mut13, is capable of improving anti-tumor activity (e.g., reducedtumor proliferation, size, volume, weight, burden or load, reduction innumber of metastatic lesions over time) of the co-administeredcheckpoint inhibitors (e.g., PD-1, PD-L1 and/or CTLA-4 or others listedherein or known in the art), e.g., by 10%, 20%, 25%, 30%, 40%, 50%, 60%,70%, 75%, 80%, 85%, 90%, 95%, or more as compared to a checkpointinhibitor therapy alone under the same conditions. In some embodiments,the anti-Galectin-9 antibody, such as any of the Galectin-9 antibodiesdescribed herein in Table 1 and/or Table 2, for example antibody 9.2-17or antibody 9.1-8mut13, is capable of improving antitumor activity(e.g., tumor proliferation, size, volume, weight, load or burden, orreduction in number of metastatic lesions over time) of theco-administered checkpoint inhibitors (e.g., PD-land/or CTLA-4 e.g.,PD-1, PD-L1 and/or CTLA-4 or others listed herein or known in the art),e.g., 1.0-1.2-fold, 1.2-1.4-fold, 1.4-1.6-fold, 1.6-1.8-fold,1.8-2-fold, or two-fold more or more as compared to a checkpointinhibitor therapy alone under the same conditions. In some embodiments,the anti-Galectin-9 antibody, such as any of the Galectin-9 antibodiesdescribed herein in Table 1 and/or Table 2, including but not limited toantibody 9.1-8m13 and/or antibody 9.2-17, is capable of improvingantitumor activity (e.g., tumor proliferation, size, volume, weight,load or burden or reduction in number of metastatic lesions over time)of the co-administered checkpoint inhibitor (e.g., PD-1, PD-L1 and/orCTLA-4 or others listed herein or known in the art), e.g., aboutthree-fold, four-fold, about threefold, four-fold, five-fold, six-fold,seven-fold, eight-fold, nine-fold, ten-fold, or more as compared to acheckpoint inhibitor therapy alone under the same conditions. In someembodiments, the co-administered checkpoint inhibitors (e.g., PD-1,PD-L1 and/or CTLA-4 or others listed herein or known in the art) arecapable of improving anti-tumor activity of the anti-Galectin-9antibody, such as any of the Galectin-9 antibodies described herein inTable 1 and/or Table 2, including but not limited to antibody 9.1-8m13and/or antibody 9.2-17, (e.g., tumor proliferation, size, volume,weight, burden or load or reduction in number of metastatic lesions),e.g., by 10%, 20%, 25%, 30%, 40%, 50%, 60%, 70%, 75%, 80%, 85%, 90%,95%, or more as compared to anti-Galectin-9 therapy alone under the sameconditions. In some embodiments, the co-administered checkpointinhibitors (e.g., PD-1, PD-L1 and/or CTLA-4 or others listed herein orknown in the art) are capable of improving antitumor activity (e.g.,tumor proliferation, size, volume, weight, load or burden or reductionin number of metastatic lesions over time) of the anti-Galectin-9antibody, such as any of the Galectin-9 antibodies described herein inTable 1 and/or Table 2, including but not limited to antibody 9.1-8m13and/or antibody 9.2-17, e.g., 1.0-1.2-fold, 1.2-1.4-fold, 1.4-1.6-fold,1.6-1.8-fold, 1.8-2-fold, or two-fold more or more as compared to ananti-Galectin-9 therapy alone under the same conditions. In someembodiments, the co-administered checkpoint inhibitors (e.g., PD-1,PD-L1 and/or CTLA-4 or others described herein or known in the art) arecapable of improving antitumor activity (e.g., tumor proliferation,size, volume, weight, load or burden or reduction in number ofmetastatic lesions over time) of the anti-Galectin-9 antibody, such asany of the Galectin-9 antibodies described herein in Table 1 and/orTable 2, including but not limited to, antibody 9.1-8m13 and/or antibody9.2-17, e.g., about three-fold, four-fold, about threefold, four-fold,five-fold, six-fold, seven-fold, eight-fold, nine-fold, ten-fold, ormore as compared to a anti-Galectin-9 therapy alone under the sameconditions.

In some embodiments, the methods are provided, wherein theanti-Galectin-9 antibody, such as any of the Galectin-9 antibodiesdescribed herein in Table 1 and/or Table 2, including but not limitedto, antibody 9.1-8m13 and/or antibody 9.2-17, is capable of improvingthe ability of the immunotherapy to activate T cells (e.g., as measuredby cytokine markers described herein) (e.g., as described herein orknown in the art), e.g., by 10%, 20%, 25%, 30%, 40%, 50%, 60%, 70%, 75%,80%, 85%, 90%, 95%, or more as compared to a immunotherapy therapy aloneunder the same conditions. In some embodiments, the anti-Galectin-9antibody is capable of improving the ability of the immunotherapy toactivate T cells (e.g., as measured by cytokine markers describedherein) (e.g., as described herein or known in the art), e.g.,1.0-1.2-fold, 1.2-1.4-fold, 1.4-1.6-fold, 1.6-1.8-fold, 1.8-2-fold, ortwo-fold more or more as compared to a immunotherapy therapy alone underthe same conditions. In some embodiments, the anti-Galectin-9 antibody,such as any of the Galectin-9 antibodies described herein in Table 1and/or Table 2, including but not limited to, antibody 9.1-8m13 and/orantibody 9.2-17, is capable of improving the ability of theimmunotherapy to activate T cells (e.g., as measured by cytokine markersdescribed herein) (e.g., as described herein or known in the art), e.g.,about three-fold, four-fold, about threefold, four-fold, five-fold,six-fold, seven-fold, eight-fold, nine-fold, ten-fold, or more ascompared to a immunotherapy therapy alone under the same conditions.

In some embodiments, the methods are provided, wherein theco-administered immunotherapies (e.g., as described herein or known inthe art) are capable of improving the ability of the anti-Galectin-9antibody, such as any of the Galectin-9 antibodies described herein inTable 1 and/or Table 2, including but not limited to, antibody 9.1-8m13and/or antibody 9.2-17, to activate T cells (e.g., as measured bycytokine markers described herein), e.g., by 10%, 20%, 25%, 30%, 40%,50%, 60%, 70%, 75%, 80%, 85%, 90%, 95%, or more as compared to ananti-Galectin-9 therapy alone under the same conditions. In someembodiments, the co-administered immunotherapies (e.g., as describedherein or known in the art) are capable of improving the ability of theanti-Galectin-9 antibody to activate T cells (e.g., as measured bycytokine markers described herein), e.g., 1.0-1.2-fold, 1.2-1.4-fold,1.4-1.6-fold, 1.6-1.8-fold, 1.8-2-fold, or two-fold more or more ascompared to an anti-Galectin-9 therapy alone under the same conditions.In some embodiments, the co-administered immunotherapies (e.g., asdescribed herein or known in the art) are capable of improving theability of the anti-Galectin-9 antibody to activate T cells (e.g., asmeasured by cytokine markers described herein), e.g., about three-fold,four-fold, about threefold, four-fold, five-fold, six-fold, seven-fold,eight-fold, nine-fold, ten-fold, or more as compared to ananti-Galectin-9 therapy alone under the same conditions.

In other embodiments, the methods are provided herein, wherein theanti-Galectin-9 antibody, such as any of the Galectin-9 antibodiesdescribed herein in Table 1 and/or Table 2, including but not limitedto, antibody 9.1-8m13 and/or antibody 9.2-17, is administered incombination with one or more of the existing modalities for treatingautoimmune disorders including, but not limited to: intravenous Igtherapy, nonsteroidal anti-inflammatory drugs (NSAID), andcorticosteroids; and anti-inflammatory treatments such as cyclosporins,rapamycins or ascomycins, or their immunosuppressive analogs, e.g.,cyclosporin A, cyclosporin G, FK-506, rapamycin,40-O-(2-hydroxy)ethyl-rapamycin etc.; cyclophosphamide; azathioprene;methotrexate; brequinar; FTY 720; leflunomide; mnizoribine; mycophenolicacid; mycophenolate mofetil; 15-deoxyspergualine; immunosuppressivemonoclonal antibodies, e.g., monoclonal antibodies to leukocytereceptors, e.g., MHC, CD2, CD3, CD4, CD7, CD25, CD28, B7, CD45, or CD58or their ligands; or other immunomodulatory compounds, e.g., CTLA4Ig, orother adhesion molecule inhibitors, e.g. mAbs or low molecular weightinhibitors including selectin antagonists and VLA-4 antagonists. Thesecombination therapies can be part of an immunomodulating regimens or aregimen for the treatment or prevention of inflammatory disorders orautoimmune disorders.

In some embodiments, the methods are provided, wherein theanti-Galectin-9 antibody, such as any of the Galectin-9 antibodiesdescribed herein in Table 1 and/or Table 2, including but not limitedto, antibody 9.1-8m13 and/or antibody 9.2-17, can also be co-used with achemotherapeutic agent, including alkylating agents, anthracyclines,cytoskeletal disruptors (Taxanes), epothilones, histone deacetylaseinhibitors, inhibitors of topoisomerase I, inhibitors of topoisomeraseII, kinase inhibitors, nucleotide analogs and precursor analogs, peptideantibiotics, platinum-based agents, retinoids, vinca alkaloids andderivatives thereof.

Non-limiting examples include: (i) anti-angiogenic agents (e.g.,TNP-470, platelet factor 4, thrombospondin-1, tissue inhibitors ofmetalloproteases (TIMP1 and TIMP2), prolactin (16-Kd fragment),angiostatin (38-Kd fragment of plasminogen), endostatin, bFGF solublereceptor, transforming growth factor beta, interferon alpha, soluble KDRand FLT-1 receptors, placental proliferin-related protein, as well asthose listed by Carmeliet and Jain (2000)); (ii) a VEGF antagonist or aVEGF receptor antagonist such as anti-VEGF antibodies, VEGF variants,soluble VEGF receptor fragments, aptamers capable of blocking VEGF orVEGFR, neutralizing anti-VEGFR antibodies, inhibitors of VEGFR tyrosinekinases and any combinations thereof; and (iii) chemotherapeuticcompounds such as, e.g., pyrimidine analogs (5-fluorouracil,floxuridine, capecitabine, gemcitabine and cytarabine), purine analogs,folate antagonists and related inhibitors (mercaptopurine, thioguanine,pentostatin and 2-chlorodeoxyadenosine (cladribine));antiproliferative/antimitotic agents including natural products such asvinca alkaloids (vinblastine, vincristine, and vinorelbine), microtubuledisruptors such as taxane (paclitaxel, docetaxel), vincristine,vinblastine, nocodazole, epothilones, and navelbine,epidipodophyllotoxins (etoposide and teniposide), DNA damaging agents(actinomycin, amsacrine, anthracyclines, bleomycin, busulfan,camptothecin, carboplatin, chlorambucil, cisplatin, cyclophosphamide,cytoxan, dactinomycin, daunorubicin, doxorubicin, epirubicin,hexamethyhnelamineoxaliplatin, iphosphamide, melphalan,merchlorehtamine, mitomycin, mitoxantrone, nitrosourea, plicamycin,procarbazine, taxol, taxotere, teniposide, triethylenethiophosphoramideand etoposide (VP16)); antibiotics such as dactinomycin (actinomycin D),daunorubicin, doxorubicin (adriamycin), idarubicin, anthracyclines,mitoxantrone, bleomycin, plicamycin (mithramycin) and mitomycin; enzymes(L-asparaginase which systemically metabolizes L-asparagine and deprivescells which do not have the capacity to synthesize their ownasparagine); antiplatelet agents; antiproliferative/antimitoticalkylating agents such as nitrogen mustards (mechlorethamine,cyclophosphamide and analogs, melphalan, chlorambucil), ethyleniminesand methylmelamines (hexamethylmelamine and thiotepa), alkylsulfonates-busulfan, nitrosoureas (carmustine (BCNU) and analogs,streptozocin), trazenes-dacarbazinine (DTIC);antiproliferative/antimitotic antimetabolites such as folic acid analogs(methotrexate); platinum coordination complexes (cisplatin,carboplatin), procarbazine, hydroxyurea, mitotane, aminoglutethimide;hormones, hormone analogs (estrogen, tamoxifen, goserelin, bicalutamide,nilutamide) and aromatase inhibitors (letrozole, anastrozole);anticoagulants (heparin, synthetic heparin salts and other inhibitors ofthrombin); fibrinolytic agents (such as tissue plasminogen activator,streptokinase and urokinase), aspirin, dipyridamole, ticlopidine,clopidogrel, abciximab; antimigratory agents; antisecretory agents(breveldin); immunosuppressives (cyclosporine, tacrolimus (FK-506),sirolimus (rapamycin), azathioprine, mycophenolate mofetil);anti-angiogenic compounds (e.g., TNP-470, genistein, bevacizumab) andgrowth factor inhibitors (e.g., fibroblast growth factor (FGF)inhibitors); angiotensin receptor blocker; nitric oxide donors;anti-sense oligonucleotides; antibodies (trastuzumab); cell cycleinhibitors and differentiation inducers (tretinoin); mTOR inhibitors,topoisomerase inhibitors (doxorubicin (adriamycin), amsacrine,camptothecin, daunorubicin, dactinomycin, eniposide, epirubicin,etoposide, idarubicin, mitoxantrone, topotecan, and irinotecan),corticosteroids (cortisone, dexamethasone, hydrocortisone,methylprednisolone, prednisone, and prednisolone); growth factor signaltransduction kinase inhibitors; mitochondrial dysfunction inducers andcaspase activators; and chromatin disruptors.

In some embodiments, methods are provided herein wherein theanti-Galectin-9 antibody, such as any of the Galectin-9 antibodiesdescribed herein in Table 1 and/or Table 2, including but not limitedto, antibody 9.1-8m13 and/or antibody 9.2-17, is administeredconcurrently with a chemotherapeutic agent. In some embodiments, methodsare provided herein, wherein the anti-Galectin-9 antibody isadministered before or after a chemotherapeutic agent. In someembodiments, methods are provided herein, wherein the chemotherapeuticagent is administered systemically. In some embodiments, methods areprovided herein, wherein the chemotherapeutic agent is administeredlocally.

In some embodiments, the methods are provided, wherein theanti-Galectin-9 antibody, such as any of the antibodies described hereinin Table 1 and/or Table 2, for example antibody 9.2-17 or antibody9.1-8mut13, is capable of improving anti-tumor activity (e.g., tumorproliferation, size, volume, weight, burden load or reduction in numberof metastatic lesions over time) of the co-administered chemotherapeuticagents (e.g., as described herein or known in the art), e.g., by 10%,20%, 25%, 30%, 40%, 50%, 60%, 70%, 75%, 80%, 85%, 90%, 95%, or more ascompared to a chemotherapeutic agent therapy alone under the sameconditions. In some embodiments, the anti-Galectin-9 antibody is capableof improving antitumor activity (e.g., tumor proliferation, size,volume, weight, load or burden or reduction in number of metastaticlesions over time) of the co-administered chemotherapeutic agents (e.g.,as described herein or known in the art), e.g., 1.0-1.2-fold,1.2-1.4-fold, 1.4-1.6-fold, 1.6-1.8-fold, 1.8-2-fold, or two-fold moreor more as compared to a chemotherapeutic agent therapy alone under thesame conditions. In some embodiments, the anti-Galectin-9 antibody iscapable of improving antitumor activity (e.g., tumor proliferation,size, volume, weight, load or burden or reduction in number ofmetastatic lesions over time) of the co-administered chemotherapeuticagent (e.g., as described herein or known in the art), e.g., aboutthree-fold, four-fold, about threefold, four-fold, five-fold, six-fold,seven-fold, eight-fold, nine-fold, ten-fold, or more as compared to achemotherapeutic agent therapy alone under the same conditions.

In some embodiments, the methods are provided, wherein theco-administered chemotherapeutic agents (e.g., as described herein orknown in the art) are capable of improving anti-tumor activity of theanti-Galectin-9 antibody, such as any of the antibodies described hereinin Table 1 and/or Table 2, for example antibody 9.2-17 or antibody9.1-8mut13, (e.g., tumor proliferation, size, volume, weight, burden orload or reduction in number of metastatic lesions over time) of, e.g.,by 10%, 20%, 25%, 30%, 40%, 50%, 60%, 70%, 75%, 80%, 85%, 90%, 95%, ormore as compared to an anti-Galectin-9 therapy alone under the sameconditions. In some embodiments, the co-administered chemotherapeuticagents (e.g., as described herein or known in the art) are capable ofimproving anti-tumor activity (e.g., tumor proliferation, size, volume,weight, burden or load or reduction in number of metastatic lesions overtime) of the anti-Galectin-9 antibody, e.g., 1.0-1.2-fold, 1.2-1.4-fold,1.4-1.6-fold, 1.6-1.8-fold, 1.8-2-fold, or two-fold more or more ascompared to an anti-Galectin-9 therapy alone under the same conditions.In some embodiments, the co-administered chemotherapeutic agents (e.g.,as described herein or known in the art) are capable of improvingantitumor activity (e.g., tumor proliferation, size, volume, weight,load or burden or reduction in number of metastatic lesions over time)of the anti-Galectin-9 antibody, e.g., about three-fold, four-fold,about threefold, four-fold, five-fold, six-fold, seven-fold, eight-fold,nine-fold, ten-fold, or more as compared to an anti-Galectin-9 therapyalone under the same conditions.

In some embodiments methods are provided herein, wherein theanti-Galectin-9 antibody, such as any of the antibodies described hereinin Table 1 and/or Table 2, for example antibody 9.2-17 or antibody9.1-8mut13, is capable of improving the ability of the chemotherapeuticagent to activate T cells (e.g., as measured by cytokine markersdescribed herein) (e.g., as described herein or known in the art), e.g.,by 10%, 20%, 25%, 30%, 40%, 50%, 60%, 70%, 75%, 80%, 85%, 90%, 95%, ormore as compared to a chemotherapeutic agent therapy alone under thesame conditions. In some embodiments, the anti-Galectin-9 antibody iscapable of improving the ability of the chemotherapeutic agent toactivate T cells (e.g., as measured by cytokine markers describedherein) (e.g., as described herein or known in the art), e.g.,1.0-1.2-fold, 1.2-1.4-fold, 1.4-1.6-fold, 1.6-1.8-fold, 1.8-2-fold, ortwo-fold more or more as compared to a chemotherapeutic agent therapyalone under the same conditions. In some embodiments, theanti-Galectin-9 antibody is capable of improving the ability of thechemotherapeutic agent to activate T cells (e.g., as measured bycytokine markers described herein) (e.g., as described herein or knownin the art), e.g., about three-fold, four-fold, about threefold,four-fold, five-fold, six-fold, seven-fold, eight-fold, nine-fold,ten-fold, or more as compared to a chemotherapeutic agent therapy aloneunder the same conditions.

In some embodiments, methods are provided herein, wherein theco-administered chemotherapeutic agents (e.g., as described herein orknown in the art) are capable of improving the ability of theanti-Galectin-9 antibody, such as any of the antibodies described hereinin Table 1 and/or Table 2, for example antibody 9.2-17 or antibody9.1-8mut13, to activate T cells (e.g., as measured by cytokine markersdescribed herein), e.g., by 10%, 20%, 25%, 30%, 40%, 50%, 60%, 70%, 75%,80%, 85%, 90%, 95%, or more as compared to an anti-Galectin-9 therapyalone under the same conditions. In some embodiments, theco-administered chemotherapeutic agents (e.g., as described herein orknown in the art) are capable of improving the ability of theanti-Galectin-9 antibody to activate T cells (e.g., as measured bycytokine markers described herein), e.g., 1.0-1.2-fold, 1.2-1.4-fold,1.4-1.6-fold, 1.6-1.8-fold, 1.8-2-fold, or two-fold more or more ascompared to an anti-Galectin-9 therapy alone under the same conditions.In some embodiments, the co-administered chemotherapeutic agents (e.g.,as described herein or known in the art) are capable of improving theability of the anti-Galectin-9 antibody to activate T cells (e.g., asmeasured by cytokine markers described herein), e.g., about three-fold,four-fold, about threefold, four-fold, five-fold, six-fold, seven-fold,eight-fold, nine-fold, ten-fold, or more as compared to ananti-Galectin-9 therapy alone under the same conditions.

In some these method embodiments, wherein the administration of ananti-Galectin-9 antibody is combined with the administration of acheckpoint inhibitor, the subject has a cancer selected from the groupconsisting of pancreatic cancer, e.g., pancreatic ductal adenocarcinoma,cholangiocarcinoma, hepatocellular carcinoma, colorectal cancer,melanoma, renal cell carcinoma, and acute myeloid leukemia. In someembodiments, methods are provided herein, wherein the anti-Galectin-9antibody is administered to a patient who is refractory to a previoustreatment, e.g., checkpoint inhibitor therapy, such as PD-1.

In some embodiments, the disclosure provides a method for treating acancer in a subject who is refractory to checkpoint inhibitor therapy,the method comprising administering to a subject in need thereof aneffective amount of an anti-Galectin-9 antibody described herein orantigen binding fragment thereof, wherein the checkpoint inhibitormolecule is selected from the group consisting of PD-1, PD-L1, PD-L2,CTLA-4, LAG3, TIM3 and A2aR. In some embodiments, the disclosureprovides a method for treating a cancer in a subject who is refractoryto checkpoint inhibitor therapy, the method comprising administering toa subject in need thereof an effective amount of an anti-Galectin-9antibody described herein or antigen binding fragment thereof, whereinthe checkpoint inhibitor molecule is PD-1. In some embodiments, thecancer is selected from the group consisting of pancreatic cancer, e.g.,pancreatic ductal adenocarcinoma, cholangiocarcinoma, hepatocellularcarcinoma, colorectal cancer, melanoma, renal cell carcinoma, and acutemyeloid leukemia. Additional useful agents can be found in, e.g.,Physician's Desk Reference, 59.sup.th edition, (2005), Thomson P D R,Montvale N.J.; Gennaro et al., Eds. Remington's The Science and Practiceof Pharmacy 20.sup.th edition, (2000), Lippincott Williams and Wilkins,Baltimore Md.; Braunwald et al., Eds. Harrison's Principles of InternalMedicine, 15.sup.th edition, (2001), McGraw Hill, NY; Berkow et al.,Eds. The Merck Manual of Diagnosis and Therapy, (1992), Merck ResearchLaboratories, Rahway N.J.

It was reported that chemotherapy and/or immune therapy of solid tumorscould enhance the level of immune modulators such as checkpointmolecules, resulting in suppressed immunity against tumor cells. Erissonet al., J. Translational Medicine (2016), 14:282; Grabosch et al., J.ImmunoTherapy of Cancer (2015), 3(suppl 2): P302; and Azad et al., EMBOJ. (2016). Anti-Galectin-9 antibodies have been found to reprogramimmune responses targeting tumor cells, particularly in PDA. As such,the co-use of an anti-Galectin-9 antibody and a chemotherapeutic agent(e.g., gemcitabine) or immunotherapeutic agent (e.g., anti-PD-L1antibody) would be expected to result in significantly enhancedtherapeutic activity against solid tumors, such as PDA.

In any of the described combination therapies, the additionaltherapeutic agent or therapy can be administered prior to,simultaneously with, or following administration of the anti-Galectin-9antibody.

Kits for Use in Treatment of Diseases Associated with Galectin-9

The present disclosure also provides kits for use in treating oralleviating a disease associated with Galectin-9, for example associatedwith Galectin-9 binding to a cell surface glycoprotein (e.g., Dectin-1,TIM3, etc.), or pathologic cells (e.g., cancer cells) expressingGalectin-9. Examples include solid tumors such as PDA and othersdescribed herein, and autoimmune diseases, such as rheumatoid arthritis,systemic lupus erythematosus (SLE), autoimmune endocrine disorders,autoimmune blood disorders, and others described herein. Such kits caninclude one or more containers comprising an anti-Galectin-9 antibody,e.g., any of those described herein, and optionally a second therapeuticagent to be co-used with the anti-Galectin-9 antibody, which is alsodescribed herein.

In some embodiments, the kit can comprise instructions for use inaccordance with any of the methods described herein. The includedinstructions can comprise a description of administration of theanti-Galectin-9 antibody, and optionally the second therapeutic agent,to treat, delay the onset, or alleviate a target disease as thosedescribed herein. The kit may further comprise a description ofselecting an individual suitable for treatment based on identifyingwhether that individual has the target disease, e.g., applying thediagnostic method as described herein. In still other embodiments, theinstructions comprise a description of administering an antibody to anindividual at risk of the target disease.

The instructions relating to the use of an anti-Galectin-9 antibodygenerally include information as to dosage, dosing schedule, and routeof administration for the intended treatment. The containers may be unitdoses, bulk packages (e.g., multi-dose packages) or subunit doses.Instructions supplied in the kits of the invention are typically writteninstructions on a label or package insert (e.g., a paper sheet includedin the kit), but machine-readable instructions (e.g., instructionscarried on a magnetic or optical storage disk) are also acceptable.

The label or package insert indicates that the composition is used fortreating, delaying the onset and/or alleviating the disease associatedwith Galectin-9 (e.g., Dectin-1 or TIM-3 signaling). Instructions may beprovided for practicing any of the methods described herein.

The kits of this invention are in suitable packaging. Suitable packagingincludes, but is not limited to, vials, bottles, jars, flexiblepackaging (e.g., sealed Mylar or plastic bags), and the like. Alsocontemplated are packages for use in combination with a specific device,such as an inhaler, nasal administration device (e.g., an atomizer) oran infusion device such as a minipump. A kit may have a sterile accessport (for example the container may be an intravenous solution bag or avial having a stopper pierceable by a hypodermic injection needle).

The container may also have a sterile access port (for example thecontainer may be an intravenous solution bag or a vial having a stopperpierceable by a hypodermic injection needle). At least one active agentin the composition is an anti-Galectin-9 antibody as those describedherein.

Kits may optionally provide additional components such as buffers andinterpretive information. Normally, the kit comprises a container and alabel or package insert(s) on or associated with the container. In someembodiments, the invention provides articles of manufacture comprisingcontents of the kits described above.

General Techniques

The practice of the present invention will employ, unless otherwiseindicated, conventional techniques of molecular biology (includingrecombinant techniques), microbiology, cell biology, biochemistry andimmunology, which are within the skill of the art. Such techniques areexplained fully in the literature, such as, Molecular Cloning: ALaboratory Manual, second edition (Sambrook, et al., 1989) Cold SpringHarbor Press; Oligonucleotide Synthesis (M. J. Gait, ed., 1984); Methodsin Molecular Biology, Humana Press; Cell Biology: A Laboratory Notebook(J. E. Cellis, ed., 1998) Academic Press; Animal Cell Culture (R. I.Freshney, ed., 1987); Introduction to Cell and Tissue Culture (J. P.Mather and P. E. Roberts, 1998) Plenum Press; Cell and Tissue Culture:Laboratory Procedures (A. Doyle, J. B. Griffiths, and D. G. Newell,eds., 1993-8) J. Wiley and Sons; Methods in Enzymology (Academic Press,Inc.); Handbook of Experimental Immunology (D. M. Weir and C. C.Blackwell, eds.); Gene Transfer Vectors for Mammalian Cells (J. M.Miller and M. P. Calos, eds., 1987); Current Protocols in MolecularBiology (F. M. Ausubel, et al., eds., 1987); PCR: The Polymerase ChainReaction, (Mullis, et al., eds., 1994); Current Protocols in Immunology(J. E. Coligan et al., eds., 1991); Short Protocols in Molecular Biology(Wiley and Sons, 1999); Immunobiology (C. A. Janeway and P. Travers,1997); Antibodies (P. Finch, 1997); Antibodies: a practical approach (D.Catty., ed., IRL Press, 1988-1989); Monoclonal antibodies: a practicalapproach (P. Shepherd and C. Dean, eds., Oxford University Press, 2000);Using antibodies: a laboratory manual (E. Harlow and D. Lane (ColdSpring Harbor Laboratory Press, 1999); The Antibodies (M. Zanetti and J.D. Capra, eds., Harwood Academic Publishers, 1995).

Without further elaboration, it is believed that one skilled in the artcan, based on the above description, utilize the present invention toits fullest extent. The following specific embodiments are, therefore,to be construed as merely illustrative, and not limitative of theremainder of the disclosure in any way whatsoever. All publicationscited herein are incorporated by reference for the purposes or subjectmatter referenced herein.

EXAMPLES Example 1: Generation of Anti-Galectin-9 Antibodies

Codon-optimized genes encoding human Galectin-9 CRD1 (residues 1-148;SEQ ID NO: 3) and mouse Galectin-9 CRD1 (residues 1-147; SEQ ID NO: 5)were cloned as GST fusions using the pGEX vector including thrombincleavage site and Avitag upstream of the cloned gene. Human Galectin-9CRD2 (residues 218-355; SEQ ID NO: 4) and mouse Galectin-9 CRD2(residues 226-353; SEQ ID NO: 6) were cloned into the pHBT vector, anIPTG inducible expression vector that contains a hexahistadine tag,Avitag and TEV cleavage site upstream of the cloned gene (Sha et al.,Proc Natl Acad Sci USA, 2013, 110: 14924-14929). Human and mouseGalectin-9 CRD2 samples were then purified via Ni-Sepharose columnsfollowed by gel filtration to apparent homogeneity and biotinylated invitro using recombinant BirA. Human and mouse Galectin-9 CRD1 sampleswere purified via GST affinity chromatography followed by thrombincleavage. Samples were further purified using gel filtrationchromatography and biotinylated in a similar manner to Galectin-9 CRD2.Recombinant full-length mouse Galectin-9 (R&D Systems) was used as acontrol where necessary.

Antibody clones capable of binding to the human or mouse Galectin-9fragments as noted above were isolated from a phage-display Fab library.The library follows the design of highly successful “Library E” (Milleret al., PloS One, 2012, 7, e43746) with improvements. A total of fourrounds of phage library sorting were performed using CRD1 and CRD2samples as the targets, essentially following published procedures(Miller et al., PloS One, 2012, 7, e43746; Fellouse et al., J Mol Biol,2007, 373, 924-940). For CRD2, selection campaigns were performed using(a) only either mouse or human CRD2 as the target or (b) using human andmouse CRD2 samples alternately in successive rounds of library sorting.For CRD1, only human CRD1 samples were used.

Binding to Galectin-9 CRDs was determined by phage ELISA (Sidhu et al.,Methods Enzymol, 2000, 328, 333-363). Biotinylated CRD samples wereimmobilized to neutravidin-coated wells and blocked with an excess ofbiotin. The wells were incubated with phage displaying single Fab clonesand then bound phages were detected with HRP-conjugated anti-M13 phageantibody.

Then, phage-displayed Fab clones were pre-incubated with 50 nMnon-biotinylated Galectin-9 CRD2 or CRD1 prior to addition to ELISAplates. Reduction in the ELISA signal of clones with competitor comparedto those without competitor indicated a high affinity and highspecificity for Galectin-9 CRD1 or CRD2.

From enriched pools of antibody clones, a total of 23 clones that bindto CRD2 (FIGS. 1A-1B) and a total of 11 clones to CRD1 (FIGS. 2A-2B)were identified. Their amino acid sequences were deduced by determiningthe DNA sequences of the Fab genes in the phage clones (which areprovided herein as SEQ ID NO: 7-75 and 77-85).

The genes for a subset of identified antibody clones were transferredinto an E. coli expression vector that has previously been described(Zhang et al., Proc Natl Acad Sci USA, 2012, 109, 8534-8539). Fabproteins were expressed in E. coli BL21 (EMD Millipore) and purifiedusing HiTrap Protein G HP column (GE Healthcare) as described (Hattoriet al., Nat Methods, 2013, 10, 992-995) followed by Superdex S200 orResourceS column (GE Healthcare). When required, purified Fab wasbiotinylated via the Avitag attached to the C-terminus of the heavychain using BirA.

Antibodies in the human IgG1, human IgG4, mouse IgG1 and mouse IgG2aformats were produced by cloning the genes for the V_(H) and V_(L)regions into mammalian expression vectors for IgG production(Invivogen). Accordingly, mIgG1 and mIgG2a are human/mouse hybrids,because the Fc (i.e. CH2 and CH2) is mouse IgG1, whereas CH1 and CL arehuman. The proteins were produced by transient transfection of ExpiCHOcells (ThermoFisher) and purified using Protein G Sepharosechromatography followed by Superdex 5200 or ResourceS chromatography (GEHealthcare).

Example 2: Characterization of Anti-Galectin-9 Antibody Clones

Epitope Binning

Whether the antibody clones bind to distinct (non-overlapping) epitopesin Galectin-9 was examined using competition phage ELISA. The binding ofall the CRD2-binding clones were significantly inhibited bypre-incubation of the purified G9.2-1, G9.2-3, G9.2-15 or G9.2-17 Fabclone (FIGS. 3A-3B), indicating that the isolated clones bind to anoverlapping epitope within CRD2. Clones G9.2-15 and G9.2-17 wereselected as representative clones for further characterization becauseof their strong binding activity and good cross-reactivity between humanand mouse Galectin-9 (FIGS. 1A-1B).

Epitope Mapping

The G9.2-17 clone was selected for further epitope analysis. Todetermine its epitope on Galectin-9 CRD2, a series of point mutants wereconstructed. Their ability to bind to G9.2-17 was assayed using phageELISA, as shown in FIG. 10A. Reductions in ELISA signal indicates siteson Galectin-9 CRD2 that are critical to G9.2-17 binding. Notably, theW309K mutation (residue numbering is according to isoform 1, NCBIGenBank Accession No. BAB83625.1) dramatically reduced the binding,while the other mutations had marginal effects, suggesting that G9.2-17binds to a region including W309. Crystal structure analysis of theregion showed that it is located opposite the sugar-binding site (FIG.10B). The term “W309” or “residue W309” refers to the tryptophan residuefound at position 309 in SEQ ID NO: 1 (Galectin-9) or to the tryptophanresidue located at position 277 in the sequence of Galectin-9 isoform 2,UniProt ID 000182-2 or to a residue in CRD2 of Galectin-9 thatcorresponds to the residue found at position 309 in SEQ ID NO: 1 or atposition 277 in the sequence of the isoform of UniProt ID 000182-2. Theterms “R253”, “R271”, “R334”, and “R341” refer to the arginine residuefound at positions 253, 271, 334, and 341, respectively, in SEQ ID NO: 1or the arginine residue found at positions 221, 239, 302, 309,respectively, in the sequence of Galectin-9 isoform 2, UniProt ID000182-2. The terms “Y330” and “Y236” refer to the tyrosine residuefound at positions 330 and 236, respectively, in SEQ ID NO: 1 or thetyrosine residue found at positions 298 and 204, respectively, in thesequence of Galectin-9 isoform 2, UniProt ID 000182-2.

Mutation that Suppresses the Interaction of G9.2-17 Fab withChromatography Matrix

Purified antibody (Fab or IgG) samples were run on TOSOH TSKgelBioassist G2WXL columns in PBS and detected using absorbance at 280 nm.The Fab sample of G9.2-17 was found to exhibit a longer retention timethan expected for its size, suggesting it interacts with thechromatography column material (FIG. 11). In comparison, the Fab sampleof G9.2-Iso eluted with the expected time. A point mutant of G9.2-17,termed G9.2.17mut6, was found to have an improved chromatography profilewhile retaining the affinity to human and mouse Galectin-9 CRD2 (FIG.12), suggesting that this mutant has a reduced level of off-targetbinding.

Antibodies that Bind to a Distinct Epitope within CRD2

Potential additional epitopes were explored using additional clones thatbind to Galectin-9 CRD2. A phage display library selection using amodified scheme so as to enrich clones that bind to an epitope that isdistinct from that of G9.2-17 was performed. Wild type humanbiotinylated Galectin-9 CRD2, the W309K Galectin-9 CRD2 mutant, orGalectin-9 CRD2 preincubated with G9.2-17 IgG was immobilized toneutravidin-coated wells and incubated with individual phage-displayedFab clones. The results are shown in FIG. 13. Three clones (G9.2-24,G9.2-25, and G9.2-26) exhibited similar levels of binding to the threetargets tested, wild-type Galectin-9 CRD2, the W309K mutant, andwild-type CRD2 in complex with G9.2-17. Their binding profiles suggestthat they bind to an epitope that is distinct from that of G9.2-17.

Affinity Measurements

The affinities of the antibodies were assessed using a bead-based assayas previously described (Nishikori et al., J Mol Biol, 2012, 424,391-399) and surface plasmon resonance (SPR). In the bead-based assay, abiotinylated protein (either a Galectin-9 sample or a Fab sample) wasimmobilized on streptavidin-coated Dynabeads M280 via thebiotin-streptavidin interaction. After blocking the excessbiotin-binding sites on the beads using unconjugated biotin, bindingtitration was performed by incubating the second component (i.e., Fabfor immobilized Galectin-9 or vice versa), followed by quantificationusing a dye-labeled neutravidin (ThermoFisher) and flow cytometryanalysis. The results obtained from this experiment are provided inFIGS. 4 and 5. In experiments where the second component is an IgG, adye-labeled anti-human IgG or anti-mouse IgG antibody was used fordetection.

In SPR experiments, a biotinylated Galectin-9 sample was immobilized onan Avicap chip (Pall ForteBio) that had been preloaded with neutravidin(ThermoFisher). Antibody samples were flowed using the OneStep method ona Pioneer SPR instrument (Pall ForteBio) and the results are provided inFIG. 6.

The above-noted two assays revealed that the analyzed Fab samples haddissociation constant (K_(D)) values in the low or sub nanomolar rangeto their respective targets, as provided in FIGS. 4-6.

Conversion of G9.2-17 into the human IgG4 format substantially reducedthe dissociation rate, as expected from the bivalent nature of IgG4(FIG. 7). This was demonstrated using the OneStep method describedabove.

Detection of Endogenous Galectin-9 on Cells

To confirm that the antibodies bind to endogenous Galectin-9 produced inhuman cells, HEK293T and CRL-2134 cell lines were incubated with abiotinylated Fab, and bound Fab was detected using neutravidinconjugated with DyLight 650. Samples were then analyzed using flowcytometry. Strong signals were observed for CRL-2134 that expressesGalectin-9 but not for HEK293T that does not express Galectin-9 (Lahm etal., J Cancer Res Clin Oncol, 2001, 127, 375-386) (FIG. 8).

The data demonstrates that these antibodies recognize endogenousGalectin-9 and also show that they have minimal cross-reactivity forother cell surface proteins. To measure antibody binding to endogenousGalectin-9 produced in human cells, CRL-2134 cells were stained usingvarying concentrations of anti-Galectin-9 antibody or a negativecontrol. Samples were then washed and bound antibodies were detectedusing anti-Mouse IgG conjugated to Dylight 650 (Invitrogen, Carlsbad,Calif.). Prior to flow cytometry analysis, propidium iodide (1 μg/mL)was added to each sample. Samples were then analyzed using flowcytometry. Percentage of galectin-9 positive cells was determined usingunstained cells as a negative control. Cell-based dissociation constant(K_(D)) was calculated from a saturation curve generated based onpercentage of galectin-9 positive cells as a function of antibodyconcentration and the 1:1 binding model

Inhibition of Galectin-9-Mediated Activation of Dectin-1 Signaling

Using a reporter cell line for human Dectin-1 signaling (Invivogen), theeffects of the anti-Galectin-9 antibodies on the signaling activationmediated by Galectin-9 were examined. In this assay, activation of theDectin-1 signaling pathway leads to the secretion of alkalinephosphatase into the cell media, which is detected as quantifiablecolorimetric changes. Cell lines were incubated with the indicatedmolecules for 16 hours. In the absence of an antibody, Galectin-9 (R&DSystems) robustly activated the reporter on par with depleted zymosan, aknown ligand for Dectin-1, as shown in FIG. 9. As expected, Galectin-9exhibited no activation on the matched cell line that did not expressDectin-1.

The antibodies inhibited the activation effect of Galectin-9, suggestingthat they block the interaction of Galectin-9 with Dectin-1 on the cellsurface.

Example 3: Evaluation of Anti-Gal-9 Antibodies in a Mouse Model ofDuctal Adenocarcinoma (PDA)

To test the effect of treatment with an anti-Gal9 antibody to pancreaticductal adenocarcinoma (PDA), two PDA mouse models can be used: theslowly progressive PDA model p48Cre;LSL-KrasG12D (KC) in which miceexpress oncogenic Kras in their pancreatic progenitor cells, and a moreaggressive orthotopic PDA model utilizing tumor cells fromPdx1Cre;LSL-KrasG12D;Tp53R172H (KPC) mice, which expresses mutant Krasand p53; as well as in human PDA42,43. A combination ofimmunohistochemical analysis, flow cytometry, or immune-fluorescentmicroscopy can be used to conduct immune profiling and assess the effectof treatment with anti-galectin antibodies as compared to isotypecontrols. Similar techniques are used to study human samples derivedfrom PDA patients.

In one example of a mouse study, six week-old KC mice are treated withthe anti-galectin antibody, e.g., G9.2-17, to test the ability ofanti-galectin antibody to reduce or prevent tumor growth. Tumorprogression is assessed, one, two, three, four, five, six, seven, andeight weeks later compared to vehicle-treated animals. Animals aresacrificed and acinar architecture in the pancreata is assessed andscored. Immune profiling is performed according to FACS methods known inthe art.

Example 4: Spheroid Preparation and Microfluidic Culture of PatientTumor Samples

Fresh tumor specimens (human patients) are received in media (DMEM) onice and minced in a 10-cm dish (on ice) using sterile forceps andscalpel. Minced tumor is resuspended in DMEM (4.5 mmol/L glucose, 100mmol/L Na pyruvate, 1:100 penicillin-streptomycin; Corning CellGro)+10%FBS (Gemini Bio-Products), 100 U/mL collagenase type IV (LifeTechnologies), and 15 mmol/L HEPES (Life Technologies). Samples arepelleted and resuspended in 10 to 20 mL media. Red blood cells (RBC) areremoved from visibly bloody samples using RBC lysis buffer (BostonBio-Products). Samples are pelleted and then resuspended in freshDMEM+10% FBS and strained over 100-μm filter and 40-μm filters togenerate 51 (>100 μm), S2 (40-100 μm), and S3 (<40 μm) spheroidfractions, which are subsequently maintained in ultralow-attachmenttissue culture plates. S2 fractions are used for ex vivo culture. Analiquot of the S2 fraction is pelleted and resuspended in type I rattail collagen (Corning) at a concentration of 2.5 mg/mL following theaddition of 10×PBS with phenol red with pH adjusted using NaOH. pH7.0-7.5 is confirmed using PANPEHA Whatman paper (Sigma-Aldrich). Thespheroid-collagen mixture is then injected into the center gel region ofa 3-D microfluidic culture device as described in Jenkins et al., CancerDiscov. 2018 February; 8(2):196-215; Ex Vivo Profiling of PD-1 BlockadeUsing Organotypic Tumor Spheroids, the contents of which is hereinincorporated by reference in its entirety. Collagen hydrogels containingpatient-derived organotypic tumor spheroids (PDOTS) are hydrated withmedia with or without anti-Gal-9 monoclonal antibodies after 30 minutesat 37° C.

In some cases, to test synergy with checkpoint inhibitors or otherimmunotherapy agents, PDOTS are treated with anti-PD-1 (pembrolizumab,250 μg/mL), anti-CTLA4 (ipilimumab, 50 μg/mL), or combination (250 μg/mLpembrolizumab+50 μg/mL ipilimumab). For indicated PDOTS studies,anti-human PD-L1 (atezolizumab at 600 μg/mL+ human IFNgamma. Immuneprofiling is performed by flow cytometry as described in Jenkins et al.

Example 5: Generation and Analysis of Variants of Clone G9.1-8

Mutants of clone G9.1-8 were designed by replacing CDR residues with Seror by truncation of the CDR regions. The mutant genes were constructedusing standard site-directed mutagenesis methods and produced asdescribed in Example 1. A total of 14 mutants were designated asG9.1-8m1-G9.1-8m1 (see Table 4-9). G9.1-8m1 and G9.1-8m2 have mutationsto CDR-H2. G9.1-8m3, G9.1-8m4 and G9.1-8m5 mutations to CDR-H3.G9.1-8m6, G9.1-8m7, G9.1-8m8, G9.1-8m8, G9.1-8m10, and G9.1-8m11 havetruncations to CDR-H3. G9.1-8m12, G9.1-8m13, and G9.1-8m14 havemutations in CDR-H2 and CDR-H3. G9.1-8m12 comprises a combination ofG9.1-8m2 and G9.1-8m8 mutations; G9.1-8m13 comprises a combination ofG9.1-8m2 and G9.1-8m9 mutations; G9.1-m14 comprises a combination ofG9.1-8m2 and G9.1-8m11 mutations.

TABLE 4 Light and Heavy Chain Sequences for G9.1-8 mutants Clone NameLight chain SEQ ID Heavy Chain SEQ ID G9.1-8m1 21 74 G9.1-8m2 21 75G9.1-8m3 21 76 G9.1-8m4 21 77 G9.1-8m5 21 78 G9.1-8m6 21 79 G9.1-8m7 2180 G9.1-8m8 21 81 G9.1-8m9 21 82 G9.1-8m10 21 83 G9.1-8m11 21 84G9.1-8m12 21 85 G9.1-8m13 21 86 G9.1-8m14 21 87

FIGS. 18-22 depict graphs showing binding data for these clones asmeasured using the bead binding assay as described in Example 2. Table5-Table XXX list the KD for various clones. As described above,antibodies mouse IgG1 and mouse IgG2a formats were produced by cloningthe genes for the V_(H) and V_(L) regions into mammalian expressionvectors for IgG production (Invivogen).

TABLE 5 Binding of purified G9.1-8 mutant mIgG1 clones as characterizedusing a bead-based binding assay Clone Designation KD (nM) G9.1-8 WTmIgG1 2.4 ± 1.1 G9.1-8m2 mIgG1 6.51 ± 0.65 G9.1-8m4 mIgG1 12.0 ± 4.2 G9.1-8m1 mIgG1 no detectable binding G9.1-8m3 mIgG1 no detectablebinding G9.1-8m5 mIgG1 no detectable binding

TABLE 6 Binding of purified G9.1-8 mutant Fab as characterized using abead-based binding assay Clone Designation KD (nM) G9.1-8 WT Fab 0.45 ±0.03 G9.1-8m6 Fab 0.52 ± 0.05 G9.1-8m7 Fab 0.93 ± 0.05 G9.1-8m8 Fab 0.56± 0.04 G9.1-8m9 Fab 1.57 ± 0.16 G9.1-8m10 Fab 58.4 ± 3.4  G9.1-8m11 Fab52.7 ± 8.8 

TABLE 7 Binding of purified G9.1-8 mutant mIgG2a as characterized usinga bead-based binding assay Clone Designation KD (nM) G9.1-8m8 mIgG2a0.51 ± 0.05 G9.1-8m9 mIgG2a 0.71 ± 0.04 G9.1-8m11 mIgG2a 1.46 ± 0.05

TABLE 8 Binding of purified G9.1-8 mutant Fab as characterized using abead-based binding assay Clone Designation KD (nM) G9.1-8 WT Fab 2.13 ±0.13 G9.1-8m12 Fab 2.48 ± 0.21 G9.1-8m13 Fab 20.7 ± 0.8  G9.1-8m14 Fab58.5 ± 7.7 

TABLE 9 Binding of purified G9.1-8 mutant mIgG2a as characterized usinga bead-based binding assay Clone Designation KD (nM) G9.1-8 WT mIgG10.63 ± 0.07 G9.1-8m12 mIgG2a 0.32 ± 0.04 G9.1-8m13 mIgG2a 0.30 ± 0.04G9.1-8m14 mIgG2a 2.01 ± 0.11

These results show that certain residues within CDR-H2 and CDR-H3 of theG9.1-8 clone can be replaced or truncated with minimal effects onantigen binding. Conversion of G9.1-8 mutant clones from the Fab intothe IgG2A format can reduce the dissociation rate, as expected from thebivalent nature of IgG (see, e.g., FIG. 21 and FIG. 22, for example,G9.1-8m13 and G9.1-8m14). Of note, with the original G9.1-8 clone andG9.1-8m8 reactivity with CRD2 was observed (FIG. 25 and data not shown).G9.1-8m12, G9.1-8m13, and G9.1-8m14 do not bind to CRD2 in a bead basedassay.

Example 6: Cellular Preparation and Flow Cytometry for Analysis of Mouseand Human Tissues

Fresh PDA tumors were placed in cold FACS buffer (PBS with 2% FBS) withCollagenase IV (1 mg/mL; Worthington Biochemical, Lakewood, N.J.),Trypsin inhibitor (1 mg/mL; EMD Millipore, Billerica, Mass.) and DNase I(2 U/mL; Promega, Madison, Wis.), and minced with scissors tosub-millimeter pieces. Tissues were then incubated at 37° C. for 20 minwith gentle shaking every 5 min. Specimens were passed through a 70 μmmesh and centrifuged at 350 g for 5 min. Cell pellets were re-suspendedin the FACS buffer and 1×106 cells were first stained with zombie yellow(BioLegend) to exclude dead cells. After viability staining, cells wereincubated with an anti-CD16/CD32 mAb (eBiosciences, San Diego, Calif.)for blocking FcγRII/II followed by antibody staining with 1 μg offluorescently conjugated extracellular mAbs. Intracellular staining forcytokines and transcription factors was performed using theFixation/Permeabilization Solution Kit (eBiosciences). For mousespecimens, we used mouse CD44 (IM7), PD-1 (29F.1A12), CD3 (17A2), CD4(RM4-5), CD8 (53-6.7), CD45 (30-F11), CD11b (M1/70), Gr1 (RB6-8C5), MHCII (M5/114.15.2), IL-10 (JESS-16E3), IFNγ (XMG1.2), TNFα (MP6-XT22),ICOS (15F9), CD69 (H1.2F3), IL-17A (TC11-18H10.1), TGFα (TW7-16B4),LFA-1 (H155-78), T-bet (eBio4B10), RORγt (AFKJS-9), and FoxP3 (FJK-16s;all eBiosciences). The antibodies were from BioLegend unless otherwisenoted. Human flow cytometry antibodies included CD45 (HI30), CD3(UCHT1), CD4 (A161A1), CD8 (HIT8a), CD44 (BJ18), TNFα (MAb11), IFNγ(4S.B3; all Biolegend). Flow cytometry was carried out on the LSR-IIflow cytometer (BD Biosciences). Data were analyzed using FlowJo v.10.1(Treestar, Ashland, Oreg.).

Example 7: Evaluation of Gal-9 Antibodies Alone or in Combination withCheckpoint Inhibition in a Mouse Model of Pancreatic Cancer and TumorMass and Immune Profile of Mice Treated with G9.2-17 mIgG1

The effect of G9.2-17 mIgG1 on tumor weight and on immune profile wasassessed in a mouse model of pancreatic cancer. 8-week old C57BL/6 male(Jackson Laboratory, Bar Harbor, Me.) mice were administeredintra-pancreatic injections of FC1242 PDA cells derived from Pdx1Cre;KrasG12D; Trp53R172H (KPC) mice (Zambirinis C P, et al., TLR9 ligationin pancreatic stellate cells promotes tumorigenesis. J Exp Med. 2015;212:2077-94). Tumor cells were suspended in PBS with 50% Matrigel (BDBiosciences, Franklin Lakes, N.J.) and 1×10⁵ tumor cells were injectedinto the body of the pancreas via laparotomy. Mice (n=10/group) receivedone pre-treatment dose i.p. followed by 3 doses (q.w.) of commercialαGalectin 9 mAb (RG9-1, 200 ug, BioXcell, Lebanon, N.H.) or G9.2-17mIgG1 (200 μg), or paired isotype, either G9.2-Iso or rat IgG2a (LTF-2,BioXcell, Lebanon, N.H.) (200 μg) (one dose per week for three weeks).Mice were sacrificed 3 weeks later and tumors were harvested foranalyses by flow cytometry. Tissue was processed and prepared and flowcytometric analysis was performed as described in Example 5.

Tumor Mass and Immune Profile of Mice Treated with G9.2-17 mIgG2a Aloneor in Combination with αPD1 mAb

The effect of G9.2-17 mIgG2a on tumor weight and on immune profile wasassessed in a mouse model of pancreatic cancer, alone or in combinationwith immunotherapy. 8-week old C57BL/6 male mice (Jackson Laboratory,Bar Harbor, Me.) were administered intra-pancreatic injections of FC1242PDA cells derived from Pdx1Cre; KrasG12D; Trp53R172H (KPC) mice. Tumorcells were suspended in PBS with 50% Matrigel (BD Biosciences, FranklinLakes, N.J.) and 1×105 tumor cells were injected into the body of thepancreas via laparotomy. Mice received one pre-treatment dose i.p.followed by 3 doses (q.w.) of G9.2-17 mIgG2a (200 μg) or a neutralizingαPD-1 mAb (29F.1A12, 200 μg, BioXcell, Lebanon, N.H.), separately or incombination, or paired isotype (LTF-2 and C1.18.4, BioXcell, Lebanon,N.H.) as indicated. Mice were sacrificed on day 26 and tumors wereharvested for analyses. Tissue was processed and prepared and flowcytometric analysis was performed as described in Example 5. Results areshown in FIGS. 17A-17C. Each point represents one mouse; *p<0.05;**p<0.01; ***p<0.001; ****p<0.0001; by unpaired Student's t-test. Theseresults show single-agent treatment with G9.2-17 mIgG2a reduces tumorgrowth at both of the dose levels, whereas anti-PD-1 alone had no effecton tumor size.

Example 8: Spheroid Preparation and Analysis of Effect of Anti-Gal9Antibody in Tumor Spheroids Derived from Patient Samples

Patient-derived organotypic tumor spheroids (PDOTS) were prepared fromfresh patient tumor specimens (pancreatic adenocarcinoma, gall bladdercancer, and liver metastasis from a colorectal cancer). Briefly,specimens were received in media on ice and minced in 10 cm dishes andresuspended in DMEM+10% FBS+100 U/mL collagenase type IV. Partiallydigested samples were pelleted, re-suspended, and strained over both 100μm and 40 μm filters to generate S1 (>100 μm), S2 (40-100 μm), and S3(<40 μm) spheroid fractions, which were subsequently maintained inlow-attachment tissue culture plates. An aliquot of the S2 fraction waspelleted and resuspended in type I rat tail collagen at a concentrationof 2.5 mg/mL following addition of 10×PBS with phenol red with pHadjusted using NaOH. The spheroid-collagen mixture was injected into thecenter gel region of the DAX-1 3D microfluidic cell culture chip. After30 minutes at 37° C., collagen hydrogels containing PDOTS were hydratedwith media and treated with Gal9 antibody (G9.2-17). Three days later,PDOTS were harvested and were flowed for immune changes. Preliminaryresults on single patient samples are shown in FIGS. 26-29. If more than100 cells were obtained, then cells were sorted for CD3+, CD4+ and CD8+,otherwise cells were only sorted for CD3+.

Example 9: Characterization of Effector Function, Cross-Reactivity andImmunogenicity of Gal-9 Antibodies

Antibody-Dependent Cell-Mediated Cytotoxicity (ADCC) Assay

In antibody-dependent cell-mediated cytotoxicity (ADCC), effector cellslyse target cells on which antibodies have bound to specific antigens onthe target cell membrane. The ADCC which is developed is designed tocharacterize the Fc effector function of antibodies and measure ADCCactivity. Although the initial assay developed relied on both target andeffector cells, the assay is further improved by directly coating theantigen of interest onto plates, bypassing the need for target cells. Arecombinant Jurkat T-cell line expressing firefly luciferase gene underthe control of NFAT response elements with constitutive expression ofhuman FcγRIIIa is used as an effector cell line. Antigen coated ontosterile ELISA plates is used as a target and the test antibody isincubated with the antigen to allow biding through the Fab fragment.Effector cells displaying the correct type of FcgR are then incubatedwith the antigen/antibody complex on the plate. When the Fc portion ofthe antibody binds to the FcγRIIIa on the surface of effector cells,receptor cross-linking leads to activation of the NFAT pathway,resulting in luciferase expression. Gene expression is allowed toproceed for 5-6 hours and luciferase activity is measured using aluminometer. A dose-response curve is generated for each antibody.

Antibody-Dependent Cell-Mediated Phagocytosis (ADCP) Assay

Antibody-dependent cell-mediated phagocytosis (ADCP) is an importantmechanism of action for antibodies that mediate part or all of theiraction though phagocytosis. In that case, antibodies mediate uptake ofspecific antigens by antigen presenting cells. ADCP can be mediated bymonocytes, macrophages, neutrophils, and dendritic cells, throughFcγRIIa, FcγRI, and FcγRIIIa, of which FcγRIIa (CD32a) on macrophagesrepresent the predominant pathway. In the ADCP assay being employed,THP-1 cells (human monocytic cell line derived from an acute monocyticleukemia patient) are used to measure ADCP. Fluorescently labeled beadsare conjugated with the target antigen, then incubated with the testantibody. THP-1 cells are then added to the plate to allow their bindingto the Fc fraction of antibodies bound to antigen-coated fluorescentbeads. Antibody binding to the beads and engagement of the Fc-receptorresults in an uptake of the beads by the THP-1 cells through aphagocytic mechanism. Phagocytosis events are analyzed using flowcytometry. Total amount of fluorescence in each cell (representing thenumber of beads phagocytosed) and the percentage offluorescence-positive cells (representing the frequency of phagocytosis)are measured. A dose-dependent curve is generated to assess the ADCPactivity mediated by each test antibody.

Cross Reactivity Assay

Off-target toxicity can present a significant problem during drugdevelopment of a therapeutic monoclonal antibody. As such, specificityis a critical factor to assess as part the characterization of any newmonoclonal candidate and an important indicator of its predicted safety.In order to assess antibody specificity and cross-reactivity, testantibodies and test samples are tested for binding against a humanproteomic array consisting of an extensive collection of humanproteins—both native and denatured—and at two working concentrations.Antibody specificity is evaluated using CDI's HuProt Human ProteomeMicroarray (˜75% of the human proteome). The microarray is incubatedwith the primary antibody, rinsed, incubated with afluorescently-labelled secondary antibody and subsequently analyzed forthe amount of fluorescence detected for each target protein. Results arecompiled as microarray images.

Detection of Anti-Drug Antibodies (ADA)

Detection of anti-drug antibodies is performed on mouse sera fromanimals treated with the anti-Gal9 mAbs in vivo. The ADA assay is run onthe Mesoscale Discovery (MSD) platform due to its increased sensitivityand dynamic range over standard ELISA methods. Biotin-conjugated Gal9and sulfo-tagged anti-Gal9 antibodies (drug) will be incubated with testsera to form a bridge complex. ADA bridging complexes will be bound tostreptavidin plates and the presence of ADA in the test serum sampleswill be detected by electrochemiluminescence detection.

Example 10: Evaluation of CRD2 Clone 17 IgG1 and IgG4 Human Galectin 9Monoclonal Antibodies in a Model of Acute Myeloid Leukemia (AML) inHumanized Mice

A study is conducted to evaluate CRD2 clone 17 IgG1 and IgG4 humanGalectin 9 monoclonal antibodies in a model of Acute Myeloid Leukemia inhumanized mice (CTG-2243, Champions). The study protocol is depicted inTable 10.

TABLE 10 AML Efficacy Study Design: Group -n- Agent Dose (ug/dose)ROA/Schedule 1 10 Vehicle Control — p.o./qwx4 2 10 Control IgG1/4 100p.o./qwx4 3 10 Control IgG1/4 200 p.o./qwx4 4 10 Control IgG 1/4 400p.o./qwx4 5 10 Anti-Gal9 1/4 100 p.o./qwx4 6 10 Anti-Gal9 1/4 200p.o./qwx4 7 10 Anti- Gal9 1/4 400 p.o./qwx4 8 10 Cytarabine  50 QDx5 910 Cytarabine vehicle — QDx5Study Animal Preparation

Animals are sublethally irradiated and reconstituted with 1-5 millionprimary AML cells via tail vein injection. In-life blood collection isperformed once monthly and flow cytometry is conducted using thefollowing flow panel: huCD45/muCD45/huCD3/huCD33 for determination ofengraftment. Once human CD33+ levels reach 20-1000 counts/ul, 6surrogate animals are euthanized for comprehensive immunophenotyping andspleen, bone marrow and peripheral blood is analyzed by the flow panelabove. Animals are randomized into treatment groups based on peripheralblood counts. Disseminated Tumor growth/burden analysis is conducted upto 42 days dosing and observation. Terminal half whole blood isprocessed and analyzed for immune parameters and serum is used for Gal9ELISA.

Terminal blood and bone marrow is collected for flow cytometry. 8-colorcell surface flow cytometry is performed from terminal bone marrow andperipheral blood from all animals: The flow panels are:LD/huCD45/huCD3/huCD33/huGalectin9/huTim9/huPD1/huCD34/huCD38/huCD117.

Fresh fecal samples are collected from all animals (1 pellet/mouse) in apolypropylene tube at baseline (prior to treatment initiation), at theend of Week1 of treatment, and at study endpoint. The collected sampleswill aree snap frozen and stored at −80° C. If possible, a terminalblood sample and tissues described is collected to assess drug toxicity.

Data Analysis

To assess animal toxicity, beginning on Day 0, animals are observeddaily and weighed 3× weekly using a digital scale; data includingindividual and mean gram weights (Mean We±SEM), mean percent weightchange versus Day 0 (% vD₀) are recorded for each group and % vD₀ isplotted at study completion. Any animal deaths are recorded daily anddesignated as drug-related (D), technical (T), tumor-related (B), orunknown (U) based on weight loss and gross observation; single agent orcombination groups reporting a mean % vD₀>20% and/or >10% mortality areconsidered above the maximum tolerated dose (MTD) for that treatment onthe evaluated regimen. Maximum mean % vD₀ (weight nadir) for eachtreatment group is reported at study completion. To assess efficacy ofthe Gal-9 antibody, tumor growth inhibition is measured. Beginning onDay 0, tumor dimensions are measured 3× weekly by digital caliper anddata, including individual and mean estimated tumor volumes (MeanTV±SEM), are recorded for each group; tumor volume (TV) is calculatedusing the formula TV=width²×length×0.52. At study completion, percenttumor growth inhibition (% TGI) values are calculated and reported foreach treatment group (T) versus control (C) using initial (i) and final(f) tumor measurements by the formula %TGI=1−(T_(f)−T_(i))/(C_(f)−C_(i)). Individual mice reporting a tumorvolume ≤30% of the Day 0 measurement for two consecutive measurementsare considered partial responders (PR). Individual mice lacking palpabletumors (0.00 mm³ for two consecutive measurements) are classified ascomplete responders (CR); a CR that persists until study completion isconsidered a tumor-free survivor (TFS). Tumor doubling time (DT) isdetermined for the vehicle treated groups using the formulaDT=(D_(f)−D_(i))*log²/(log TV_(f)−log TV_(i)) where D=Day and TV=TumorVolume. All data collected in this study is managed electronically andstored on a redundant server system.

Example 11: Evaluation of Gal-9 Antibody in a B16F10 Melanoma SyngeneicTumor Model in Immunocompetent Mice

Gal-9 antibody G9.2-17 was evaluated in the B16F10 syngeneic mouse modelof melanoma immunocompetent mice. Pre-study animals (female C57BL/6, 6-8weeks of age (Charles River Labs)) were unilaterally implantedsubcutaneously on the left flank with 5e5 B16.F10 in 100 μl PBS.Pre-study tumor volumes were recorded for each experiment beginning 2-3days after implantation. When tumors reached an average tumor volume of50-100 mm³ (preferably 50-75 mm³) animals were matched by tumor volumeinto treatment or control groups (n=8) to be used for dosing and dosingwas initiated on Day 0. Animals were dosed on day 0 and day 4 i.v. Thestudy design for testing of Anti-Gal9 G9.2-17 IgG1 and Anti-Gal9 G9.2-17IgG2 is summarized in Table 11 and Table 12.

TABLE 11 Anti-Gal9 IgG1 Route of Dose Dose Administration Group -n- TestAgent (ug/mouse) Volume (ROA) 1 8 Control Untreated — — — 2 8 ControlmIgG1 200 ug 200 ul IV 7 8 Anti-Gal9 mIgG1 200 ug 200 ul IV (G9.2-17)

TABLE 12 Anti-Gal9 IgG2 Route of Total Dose Dose Administration NumberGroup -n- Test Agent (ug/mouse) Volume (ROA) Schedule of Doses 1 10Control Untreated — — — — — 2 10 Control mIgG2 200 ug 200 ul IV Q4Dx6 63 10 Control mIgG2 200 ug 200 ul IP BIWx4 8 4 10 Anti-Gal9 mIgG2 200 ug200 ul IV Q4Dx6 6 (G9.2-17)

Tumor volumes were taken and animals were weighed three times weekly.The study endpoint was set when the mean tumor volume of the controlgroup (uncensored) reached 1500 mm3. A final tumor volume was taken onthe day the study reached endpoint. A final weight was taken on the daythe study reached end point (day 10). Tumor volume is shown in FIG. 30and FIG. 31.

Example 12: Jurkat Apoptosis Assay

The ability of Galectin-9 G9.2-17 and G9.1-8m9 antibodies to preventGalectin-9-induced apoptosis of Jurkat cells was assessed. Jurkat cells(TIB-152, ATCC, Manassas, Va.) were grown in modified RPMI (2 mML-glutamine, 10 mM HEPES, 1 mM sodium pyruvate, 4.5 g/L glucose, 1.5 g/Lsodium bicarbonate) with 10% FBS, 100 mU/mL penicillin and 100 μg/mLstreptomycin at 37° C. with 5% CO₂. Cells (2×10⁵ cells/well) wereincubated in the wells of a 96-well culture plate with or without theaddition of 280 nM Galectin-9 (2045-GA, RnD Systems, Minneapolis,Minn.), 1 μM G9.2-17 IgG, and/or 1 μM G9.1-8m9 IgG. Cells were incubatedat 37° C., 5% CO₂ for 5 hours and then resuspended in annexinV-bindingbuffer. Cells were then stained with AnnexinV-AlexaFluor488 (Invitrogen,Carlsbad, Calif.) at 4° C. for 30 minutes in the dark. Prior to flowcytometry, propidium iodide (PI) was added (1 μg/mL). Cells were run onGuava easyCyte flow cytometer (MilliporeSigma, Burlington, Mass.) andanalyzed using FlowJo (Treestar, Ashland, Oreg.). Cell population wasgated via forward and side scatter, then analyzed on AnnexinV for allapoptotic cells and PI for late apoptotic cell populations. Results areshown in FIG. 23.

Example 13. Anti-Galectin-9 Antibodies Disrupt the Interaction BetweenGalectin-9 and CD206

Microlon high binding 96 well plates (Greiner Bio-One, Kremsmünster,Austria) were coated with hGalectin-9 (RnD Systems, Minneapolis, Minn.)(50 μL, 4 μg/mL in TBS) at room temperature for 1 hour. Plates were thenblocked with TBS+0.5% BSA (150 μL) at room temperature for 1 hour. Wellswere washed three times with TBS+0.1% BSA. G9.1-8m13, and/or G9.2-17antibodies were added to each well (50 μL, 100 nM in TBS+0.1% BSA) andincubated at room temperature for 30 minutes. CD206-His (RnD Systems,Minneapolis, Minn.) was then spiked in to a final concentration of 13 nMand incubated for an additional 30 minutes at room temperature. Wellswere then washed 3 times with TBS+0.05% Tween20. aHis-HRP (ab1187,Abcam, Cambridge, Mass.) was added to each well (50 μL, 1:2500 inTBS+0.05% Tween20, 0.1% BSA) and incubated an additional 30 minutes atroom temperature. Wells were washed three times with TBS+0.05% Tween20and once with TBS. 1Step TMB-Ultra ELISA substrate (Thermofisher,Waltham, Mass.) (50 μL) was added to each well.

The reaction was neutralized with 2M H2504 (50 μL) and absorbance signalat 450 nm was read using an Epoch2 spectrophotometer (BioTek, Winooski,Vt.). Experiments were performed in triplicate; *p<0.05; **p<0.01;***p<0.001; ****p<0.0001; by unpaired Student's t-test). Results areshown in FIG. 24A and FIG. 24B and indicate that both G9.1-8m13 andG9.2-17 antibodies inhibit the interaction between Galectin-9 and CD206and their effects are additive.

Example 14. Evaluation of Cytotoxic Activity

To evaluate complement-induced cytotoxic activity of 9.1-8mut13 and9.2-17, complement-dependent cytotoxicity (CDC) assays are performed andcompared with 2 mouse monoclonal antibodies (mouse monoclonal forms ofg9.2-17 and g9.1-8m9). Antibodies are incubated with the appropriatetarget cells expressing either gal-9, and species-specific serum isadded as a source of complement proteins to bind to the cell-boundmonoclonal antibodies and initiate complement-dependent cytolysis of thetarget cell. Cell death of target cells is determined by thedifferential staining obtained in cells with permeable vs. non-permeablemembranes (i.e., lysed vs. unlysed cells) after incubation with afluorescent cell viability dye and assessed by flow cytometry.

Example 15: Evaluation of Gal-9 Antibody in Two Syngeneic Models ofColorectal and Melanoma Cancer in Immunocompetent Mice

Gal-9 antibodies G9.2-17 and G9.1-8m13 are evaluated in syngeneic modelsof colorectal and melanoma cancer in immunocompetent mice. Test articlesare formulated and prepared on a weekly basis for the duration of thestudy according to Table 13.

TABLE 13 Test articles Master Master Working Working Stock Stock StockStock Agent Storage Stability State Storage Stability Control   4° C.,For the Liquid   4° C., For the mIgG1 Dark duration Dark duration ofstudy of study Control   4° C., For the Liquid   4° C., For the mIgG2Dark duration Dark duration of study of study Gal9-IgG1 −80° C. For theLiquid −20° C. For the (G9.2-17) duration duration of study of studyGal9-IgG2 −80° C. For the Liquid −20° C. For the (G9.2-17) durationduration of study of study Gal9-IgG1 −80° C. For the Liquid −20° C. Forthe (G9.1-8m13) duration duration of study of study mGal9-IgG2 −80° C.For the Liquid −20° C. For the (G9.1-8m13) duration duration of study ofstudy anti-mPD-1   4° C., For the Liquid   4° C., For the Dark durationDark duration of study of studyVehicle Control: mGal9-IgG1, and mGal9-IgG2; Control mIgG1, ControlmIgG2, and anti-mPD-1: Sterile PBSExperimental Design

Pre-study animals (female C57BL/6, 6-8 weeks of age (Charles River Labs)are acclimatized for 3 days and then are unilaterally implantedsubcutaneously on the left flank with 5e5 B16.F10 (melanoma cell line)or MC38 cells (colorectal cancer cell line) resuspended in 100 μl PBS.Pre-study tumor volumes are recorded for each experiment beginning 2-3days after implantation. When tumors reach an average tumor volume of50-100 mm³ (preferably 50-75 mm³) animals are matched by tumor volumeinto treatment or control groups to be used for dosing and dosinginitiated on Day 0. The study design for testing of Anti-Gal9 IgG1 andAnti-Gal9 IgG2 is summarized in Table 14 and Table 15.

TABLE 14 Anti-Gal9 IgG1 (B16F10 and MC38) Route of Total Dose DoseAdministration Number Group -n- Test Agent (ug/mouse) Volume (ROA)Schedule of Doses 1 8 Control Untreated — — — — — 2 8 Control mIgG1 200ug 200 ul IV Q4Dx6 6 3 8 Control mIgG1 400 ug 200 ul IV Q4Dx6 6 4 8Control mIgG2 200 ug 200 ul IP BIWx4 8 5 8 Anti-Gal9 mIgG1 200 ug 200 ulIV Q4Dx6 6 6 8 Anti-Gal9 mIgG1 400 ug 200 ul IV Q4Dx6 6 7 8 Anti-Gal9mIgG1 200 ug 200 ul IV Q4Dx6 6 (G9.1-8m13) 8 8 Anti-Gal9 mIgG1 400 ug200 ul IV Q4Dx6 6 (G9.1-8m13) 9 8 Anti-Gal9 mIgG1 + 200 ug 200 ul IVQ4Dx6 6 mAnti-PD1 200 ug 200 ul IP BIWx4 8 10 8 Anti-Gal9 mIgG1 + 400 ug200 ul IV Q4Dx6 6 mAnti-PD1 200 ug 200 ul IP BIWx4 8 11 8 Anti-Gal9mIgG1 200 ug 200 ul IV Q4Dx6 6 (G9.1-8m13) + 200 ug 200 ul IP BIWx4 8mAnti-PD1 12 8 Anti-Gal9 mIgG1 400 ug 200 ul IV Q4Dx6 6 (G9.1-8m13) +200 ug 200 ul IP BIWx4 8 mAnti-PD1 13 8 mAnti-PD1 200 ug 200 ul IP BIWx48

TABLE 15 Anti-Gal9 IgG2 (B16F10 and MC38) Route of Total Dose DoseAdministration Number Group -n- Test Agent (ug/mouse) Volume (ROA)Schedule of Doses 1 10 Control Untreated — — — — — 2 10 Control mIgG2200 ug 200 ul IV Q4Dx6 6 3 10 Control mIgG2 400 ug 200 ul IV Q4Dx6 6 410 Control mIgG2 200 ug 200 ul IP BIWx4 8 5 10 Anti-Gal9 mIgG2 200 ug200 ul IV Q4Dx6 6 6 10 Anti-Gal9 mIgG2 400 ug 200 ul IV Q4Dx6 6 5 10Anti-Gal9 mIgG2 200 ug 200 ul IV Q4Dx6 6 (G9.1-8m13) 6 10 Anti-Gal9mIgG2 400 ug 200 ul IV Q4Dx6 6 (G9.1-8m13) 7 10 Anti-Gal9 mIgG2 + 200 ug200 ul IV Q4Dx6 6 mAnti-PD1 200 ug 200 ul IP BIWx4 8 8 10 Anti-Gal9mIgG2 + 400 ug 200 ul IV Q4Dx6 6 mAnti-PD1 200 ug 200 ul IP BIWx4 8 7 10Anti-Gal9 mIgG2 200 ug 200 ul IV Q4Dx6 6 (G9.1-8m13) + 200 ug 200 ul IPBIWx4 8 mAnti-PD1 8 10 Anti-Gal9 mIgG2 400 ug 200 ul IV Q4Dx6 6(G9.1-8m13) + 200 ug 200 ul IP BIWx4 8 mAnti-PD1 9 10 mAnti-PD1 200 ug200 ul IP BIWx4 8

Tumor volumes are taken three times weekly. A final tumor volume istaken on the day the study reaches endpoint. A final tumor volume istaken if an animal is found moribund. Animals are weighed three timesweekly. A final weight is taken on the day the study reaches end pointor if animal is found moribund. Animals exhibiting ≥10% weight loss whencompared to Day 0 are provided DietGel® ad libitum. Any animalexhibiting >20% net weight loss for a period lasting 7 days or if micedisplay >30% net weight loss when compared to Day 0 is consideredmoribund and is euthanized. The study endpoint is set when the meantumor volume of the control group (uncensored) reaches 1500 mm3. If thisoccurs before Day 28, treatment groups and individual mice are dosed andmeasured up to Day 28. If the mean tumor volume of the control group(uncensored) does not reach 1500 mm3 by Day 28, then the endpoint forall animals is the day when the mean tumor volume of the control group(uncensored) reaches 1500 mm3 up to a maximum of Day 60. Blood iscollected from all animals from each group. For blood collection, asmuch blood as possible is collected via a cardiac puncture into K₂EDTAtubes (400 ul) and serum separator tubes (remaining) under deepanesthesia induced by isoflurane inhalation. The blood collected intoK₂EDTA tubes is placed on wet ice until used for performing immune panelflow as shown in Table 16.

TABLE 16 Flow Cytometry Panel 1 Antibody Description Conjugate CloneSupplier mCD3 FITC 17A2 BioLegend mCD4 APC-Fire RM4-4 BioLegend 750mGamma BV605 GL3 BioLegend Delta mCD8 APC-R700 53-6.7 BioLegend mCD44BV786 IM7 BioLegend mCD11b APC M1/70 BioLegend mCD45 BV510 30-F11BioLegend Live Dead 7AAD — BioLegend mCD62L PE-Cy7 MEL-14 BioLegendmPD-1 BV711 29F.1A12 BioLegend mCTLA4 PE UC10-4B9 BioLegend mCD27 BV421LG.3A10 BioLegend

Blood collected into serum separator tubes is allowed to clot at roomtemperature for at least 15 minutes. Samples are centrifuged at 3500 for10 minutes at room temperature. The resultant serum is separated,transferred to uniquely labeled clear polypropylene tubes, and frozenimmediately over dry ice or in a freezer set to maintain −80° C. untilshipment for the bridging ADA assay (shipped within one week).

Tumors from all animals are collected as follows. Tumors less than 400mm³ in size are snap frozen, placed on dry ice, and stored at −80C untilused for RT-qPCR analysis. For tumors of 400-500 mm³ in size, wholetumors are collected into MACS media for use in the Flow Panel (shown inTable 16A below). For tumors greater than 500 mm³ in size, a small piece(about 50 mm³) is snap frozen placed on dry ice, and stored at −80C forRT-qPCR, and the remaining tumor is collected in MACS media for flowcytometry (as shown in Table 16A). For flow cytometry, tumors are placedin MACS media and stored on wet ice until processed. A summary of theflow cytometry analysis performed is shown in Table 16A.

TABLE 16A Flow cytometry Panel 2 Antibody Description Conjugate CloneSupplier mCD3 FITC 17A2 BioLegend mCD4 APC-Fire RM4-4 BioLegend 750mGamma BV605 GL3 BioLegend Delta mCD8 APC-R700 53-6.7 BioLegend mCD69BV421 H1.2F3 BioLegend mCD11b APC M1/70 BioLegend mCD45 BV510 30-F11BioLegend Live Dead 7AAD — BioLegend mCD62L PE-Cy7 MEL-14 BioLegendmPD-1 BV711 29F.1A12 BioLegend mCTLA4 PE UC10-4B9 BioLegend mNk1.1 BV786PK136 BioLegend

Spleen, liver, colon, lungs, heart, and kidneys from all animals areretained in 10% neutral buffered formalin (NBF) for 18-24 hours,transferred to 70% ethanol and stored at room temperature. Formalinfixed samples are paraffin embedded.

Example 16: Evaluation of Gal-9 Antibody in a Models ofCholangiocarcinoma

The efficacy of Gal-9 antibody is assessed in a mouse model ofcholangiocarcinoma as described in S. Rizvi, et al. (YAP-associatedchromosomal instability and cholangiocarcinoma in mice, Oncotarget, 9(2018) 5892-5905), the contents of which is herein incorporated byreference in its entirety. In this transduction model, in whichoncogenes (AKT/YAP) are instilled directly into the biliary tree, tumorsarise from the biliary tract in immunocompetent hosts withspecies-matched tumor microenvironment. Dosing is described in Table 17.

TABLE 17 Dosing Route of Total Dose Dose Administration Number of Group-n- Test Agent (ug/mouse) Volume (ROA) Schedule Doses 1 10 ControlUntreated — — — — — 2 10 Control mIgG2 200 ug 200 ul IV Q4Dx6 6 3 10Control mIgG2 400 ug 200 ul IV Q4Dx6 6 4 10 Control mIgG2 200 ug 200 ulIP BIWx4 8 5 10 Anti-Gal9 mIgG2 200 ug 200 ul IV Q4Dx6 6 (G9.2-17) 6 10Anti-Gal9 mIgG2 400 ug 200 ul IV Q4Dx6 6 (G9.2-17) 7 10 Anti-Gal9 mIgG2200 ug 200 ul IV Q4Dx6 6 (G9.1.8-m13) 8 10 Anti-Gal9 mIgG2 400 ug 200 ulIV Q4Dx6 6 (G9.1.8-m13)

In brief, murine CCA cells (described in S. Rizvi, et al) are harvestedand washed in DMEM. Male C57BL/6 mice from Jackson Labs are anesthetizedusing 1.5-3% isoflurane. Under deep anesthesia, the abdominal cavity isopened by a 1 cm incision below the xiphoid process. A sterile cottontipped applicator is used to expose the superolateral aspect of themedial lobe of the liver. Using a 27-gauge needle, 40 μL of standardmedia containing 1×10^6 cells is injected into the lateral aspect of themedial lobe. Cotton tipped applicator is held over the injection site toprevent cell leakage and blood loss. Subsequently, the abdominal walland skin are closed in separate layers with absorbable chromic 3-0 gutsuture material.

Two weeks post implantation, animals are matched by tumor volume intotreatment or control groups to be used for dosing and dosing initiatedon Day 0. Tumor volumes are measured and animals weighed three timesweekly. A final tumor volume and weight is taken on the day the studyreaches endpoint (4 weeks or when tumor burden of control becomes 1500mm3). Blood is collected from all animals from each group. Tumors fromall animals are collected essentially as described in Example 15.Analysis is performed essentially as described in Example 15.

EQUIVALENTS

From the above description, one skilled in the art can easily ascertainthe essential characteristics of the present invention, and withoutdeparting from the spirit and scope thereof, can make various changesand modifications of the invention to adapt it to various usages andconditions. Thus, other embodiments are also within the claims.

While several inventive embodiments have been described and illustratedherein, those of ordinary skill in the art will readily envision avariety of other means and/or structures for performing the functionand/or obtaining the results and/or one or more of the advantagesdescribed herein, and each of such variations and/or modifications isdeemed to be within the scope of the inventive embodiments describedherein. More generally, those skilled in the art will readily appreciatethat all parameters, dimensions, materials, and configurations describedherein are meant to be exemplary and that the actual parameters,dimensions, materials, and/or configurations will depend upon thespecific application or applications for which the inventive teachingsis/are used. Those skilled in the art will recognize, or be able toascertain using no more than routine experimentation, many equivalentsto the specific inventive embodiments described herein. It is,therefore, to be understood that the foregoing embodiments are presentedby way of example only and that, within the scope of the appended claimsand equivalents thereto, inventive embodiments may be practicedotherwise than as specifically described and claimed. Inventiveembodiments of the present disclosure are directed to each individualfeature, system, article, material, kit, and/or method described herein.In addition, any combination of two or more such features, systems,articles, materials, kits, and/or methods, if such features, systems,articles, materials, kits, and/or methods are not mutually inconsistent,is included within the inventive scope of the present disclosure.

All definitions, as defined and used herein, should be understood tocontrol over dictionary definitions, definitions in documentsincorporated by reference, and/or ordinary meanings of the definedterms.

All references, patents and patent applications disclosed herein areincorporated by reference with respect to the subject matter for whicheach is cited, which in some cases may encompass the entirety of thedocument.

The indefinite articles “a” and “an,” as used herein in thespecification and in the claims, unless clearly indicated to thecontrary, should be understood to mean “at least one.”

The phrase “and/or,” as used herein in the specification and in theclaims, should be understood to mean “either or both” of the elements soconjoined, i.e., elements that are conjunctively present in some casesand disjunctively present in other cases. Multiple elements listed with“and/or” should be construed in the same fashion, i.e., “one or more” ofthe elements so conjoined. Other elements may optionally be presentother than the elements specifically identified by the “and/or” clause,whether related or unrelated to those elements specifically identified.Thus, as a non-limiting example, a reference to “A and/or B”, when usedin conjunction with open-ended language such as “comprising” can refer,in one embodiment, to A only (optionally including elements other thanB); in another embodiment, to B only (optionally including elementsother than A); in yet another embodiment, to both A and B (optionallyincluding other elements); etc.

As used herein in the specification and in the claims, “or” should beunderstood to have the same meaning as “and/or” as defined above. Forexample, when separating items in a list, “or” or “and/or” shall beinterpreted as being inclusive, i.e., the inclusion of at least one, butalso including more than one, of a number or list of elements, and,optionally, additional unlisted items. Only terms clearly indicated tothe contrary, such as “only one of” or “exactly one of,” or, when usedin the claims, “consisting of,” will refer to the inclusion of exactlyone element of a number or list of elements. In general, the term “or”as used herein shall only be interpreted as indicating exclusivealternatives (i.e., “one or the other but not both”) when preceded byterms of exclusivity, such as “either,” “one of,” “only one of,” or“exactly one of.” “Consisting essentially of,” when used in the claims,shall have its ordinary meaning as used in the field of patent law.

As used herein in the specification and in the claims, the phrase “atleast one,” in reference to a list of one or more elements, should beunderstood to mean at least one element selected from any one or more ofthe elements in the list of elements, but not necessarily including atleast one of each and every element specifically listed within the listof elements and not excluding any combinations of elements in the listof elements. This definition also allows that elements may optionally bepresent other than the elements specifically identified within the listof elements to which the phrase “at least one” refers, whether relatedor unrelated to those elements specifically identified. Thus, as anon-limiting example, “at least one of A and B” (or, equivalently, “atleast one of A or B,” or, equivalently “at least one of A and/or B”) canrefer, in one embodiment, to at least one, optionally including morethan one, A, with no B present (and optionally including elements otherthan B); in another embodiment, to at least one, optionally includingmore than one, B, with no A present (and optionally including elementsother than A); in yet another embodiment, to at least one, optionallyincluding more than one, A, and at least one, optionally including morethan one, B (and optionally including other elements); etc.

It should also be understood that, unless clearly indicated to thecontrary, in any methods claimed herein that include more than one stepor act, the order of the steps or acts of the method is not necessarilylimited to the order in which the steps or acts of the method arerecited.

What is claimed is:
 1. An isolated antibody, which binds humangalactin-9, wherein the antibody comprises a heavy chain complementaritydetermining region 1 (CDR1) set forth as SEQ ID NO: 361, a heavy chaincomplementary determining region 2 (CDR2) set forth as SEQ ID NO: 388,and a heavy chain complementary determining region 3 (CDR3) set forth asSEQ ID NO: 406 and/or comprises a light chain complementaritydetermining region 1 (CDR1) set forth as SEQ ID NO: 328, a light chaincomplementary determining region 2 (CDR2) set forth as SEQ ID NO: 329,and a light chain complementary determining region 3 (CDR3) set forth asSEQ ID NO:
 352. 2. The isolated antibody of claim 1, wherein theantibody binds to an epitope in a carbohydrate recognition domain (CRD)of galectin-9.
 3. The isolated antibody of claim 2, wherein the CRD isCRD2.
 4. The isolated antibody of claim 3, wherein the CRD2 consists ofthe amino acid sequence of SEQ ID NO:
 4. 5. The isolated antibody ofclaim 1, which is a full-length antibody or an antigen-binding fragmentthereof.
 6. The isolated antibody of claim 1, which is a single chainantibody.
 7. The isolated antibody of claim 1, which is a human antibodyor a humanized antibody.
 8. The isolated antibody of claim 1, whereinthe antibody comprises a VH set forth as SEQ ID NO: 55 and a VL setforth as SEQ ID NO:
 54. 9. The isolated antibody of claim 8, wherein theantibody binds to an epitope in a carbohydrate recognition domain (CRD)of galectin-9, wherein the CRD is CRD2.
 10. The isolated antibody ofclaim 8, which is a full-length antibody or an antigen-binding fragmentthereof.
 11. The isolated antibody of claim 8, which is a single chainantibody.
 12. The isolated antibody of claim 8, which is a humanantibody or a humanized antibody.
 13. The isolated antibody of claim 8,which is an IgG molecule.
 14. The isolated antibody of claim 13, whereinthe antibody is an IgG1 or IgG4 molecule.
 15. The isolated antibody ofclaim 14, wherein the antibody is an IgG4 molecule, wherein the IgG4 hasa S228P mutation.
 16. The isolated antibody of claim 14, wherein theantibody comprises a heavy chain constant region set forth as SEQ ID NO:422, and/or a light chain constant region set forth as SEQ ID NO: 418.17. The isolated antibody of claim 8, wherein the antibody comprises aV_(H) set forth as SEQ ID NO: 55 and a V_(L) set forth as SEQ ID NO: 54,and has a heavy chain constant region set forth as SEQ ID NO: 422 and alight chain constant region set forth as SEQ ID NO:
 418. 18. Theisolated antibody of claim 17, wherein the antibody comprises a heavychain set forth as SEQ ID NO: 316 and a light chain set forth as SEQ IDNO:108.
 19. The isolated antibody of claim 18, wherein the antibody isG9.2-17.
 20. The isolated antibody of claim 19, wherein the antibodybinds to an epitope in a carbohydrate recognition domain (CRD) ofgalectin-9, wherein the CRD is CRD2.
 21. A pharmaceutical compositioncomprising the antibody of claim 19 and a pharmaceutically acceptablecarrier.
 22. A pharmaceutical composition comprising the antibody ofclaim 17 and a pharmaceutically acceptable carrier.
 23. A pharmaceuticalcomposition comprising the antibody of claim 8 and a pharmaceuticallyacceptable carrier.
 24. The isolated antibody of claim 1, which is anIgG molecule.
 25. The isolated antibody of claim 24, wherein theantibody is an IgG1 or IgG4 molecule.
 26. The isolated antibody of claim25, wherein the antibody is an IgG4 molecule, wherein the IgG4 has aS228P mutation.
 27. The isolated antibody of claim 25, wherein theantibody comprises a heavy chain constant region set forth as SEQ ID NO:422, and/or a light chain constant region set forth as SEQ ID NO: 418.28. The isolated antibody of claim 1, wherein the antibody has a V_(L)CDR1 set forth as SEQ ID NO: 328, a V_(L) CDR2 set forth as SEQ ID NO:329, a V_(L) CDR3 set forth as SEQ ID NO: 352, a V_(H) CDR1 set forth asSEQ ID NO: 361, a V_(H) CDR2 set forth as SEQ ID NO: 388, a V_(H) CDR3set forth as SEQ ID NO: 406, and has a heavy chain constant region setforth as SEQ ID NO: 422 and a light chain constant region set forth asSEQ ID NO:
 418. 29. The isolated antibody of claim 1, wherein theantibody comprises a heavy chain set forth as SEQ ID NO: 316 and/or alight chain set forth as SEQ ID NO:108.
 30. A pharmaceutical compositioncomprising the antibody of claim 1 and a pharmaceutically acceptablecarrier.